Perspectives of the International Society of Cell & Gene Therapy Gastrointestinal Scientific Committee on the Intravenous Use of Mesenchymal Stromal Cells in Inflammatory Bowel Disease (PeMeGi)
Publication date: Available online 11 June 2019Source: CytotherapyAuthor(s): R. Ciccocioppo, D.C. Baumgart, C.C. Dos santos, J. Galipeau, C. Klersy, G. OrlandoAbstractInflammatory bowel disease (IBD), namely, Crohn's disease and ulcerative colitis, remains a grievous and recalcitrant problem incurring significant human and health care costs, even in consideration of the growing incidence. Initial goals of care aimed to achieve the induction and maintenance of clinical remission. The advent of novel treat-to-target approaches using patient stratification, early introduction of immunosuppressants and rapid escalation to biol...
Source: Cytotherapy - June 12, 2019 Category: Cytology Source Type: research

Current state of Health Canada regulation for cellular and gene therapy products: potential cures on the horizon
Publication date: Available online 10 June 2019Source: CytotherapyAuthor(s): JOLENE CHISHOLM, CRYSTAL RUFF, SOWMYA VISWANATHANAbstractWe provide an overview of the regulatory framework, pathways and underlying regulatory authority for cell, gene and tissue-engineered therapies in Canada. Canada's regulatory approach uses three sets of regulations, namely, the Cells, Tissues and Organs Regulations, the Food and Drug Regulations and the Medical Devices Regulations. We provide an overview of each these sets of regulations as they apply to clinical investigation to post-market product lifecycle stages. Information is provided ...
Source: Cytotherapy - June 11, 2019 Category: Cytology Source Type: research

Current state of U.S. Food and Drug Administration regulation for cellular and gene therapy products: potential cures on the horizon
Publication date: Available online 10 June 2019Source: CytotherapyAuthor(s): MICHAEL Mendicino, YONG FAN, DEBORAH GRIFFIN, KURT C. GUNTER, KAREN NICHOLSAbstractCellular & Gene Therapies (CGTs) are complex products, which have been key foci of the International Society for Cell & Gene Therapy (ISCT). For this ISCT North American Legal & Regulatory Affairs Committee review publication, CGTs include but are not limited to somatic cell-based therapies, pluripotent cell-derived cell-based therapies, gene- or non-gene-modified or gene edited versions of these cell-based therapies, in vivo gene therapies, organ/tissue...
Source: Cytotherapy - June 11, 2019 Category: Cytology Source Type: research

Microporous acellular extracellular matrix combined with adipose-derived stem cell sheets as a promising tissue patch promoting articular cartilage regeneration and interface integration
Publication date: Available online 11 June 2019Source: CytotherapyAuthor(s): Yuxiang Zhang, Gang Feng, Guoping Xu, Yiying QiAbstractBackgroundAcute or chronic injury of articular cartilage leads to localized destruction. Difficulties with interface integration between the implant and native cartilage tissue can lead to an undesirable outcome. To improve cartilage repair and interface integration, we explored the therapeutic efficacy of microporous acellular extracellular matrix (ECM) combined with adipose-derived stem cell (ASC) sheets.MethodsMethods for fabricating ASC sheets and microporous acellular ECM were explored be...
Source: Cytotherapy - June 11, 2019 Category: Cytology Source Type: research

Manufacturing mesenchymal stromal cells for clinical applications: A survey of Good Manufacturing Practices at U.S. academic centers
ConclusionsSurvey responses reported herein offer insight into the current best practices used to manufacture MSC-based products in the United States and how these practices may affect product quality and potency. The responses also provide a foundation to establish standardized manufacturing platforms. (Source: Cytotherapy)
Source: Cytotherapy - June 8, 2019 Category: Cytology Source Type: research

The efficacy of anti-CD19 chimeric antigen receptor T cells for B-cell malignancies
In conclusion, this meta-analysis showed a high clinical RR of CD19-CAR-T cell–based immunotherapy in patients with refractory B-cell malignancies. (Source: Cytotherapy)
Source: Cytotherapy - June 1, 2019 Category: Cytology Source Type: research

A highly standardized and characterized human platelet lysate for efficient and reproducible expansion of human bone marrow mesenchymal stromal cells
ConclusionsWe report a standardization of hPL and the importance of such standardization for the efficient amplification of more homogeneous and reproducible cell therapy products. (Source: Cytotherapy)
Source: Cytotherapy - May 26, 2019 Category: Cytology Source Type: research

Identification of senescent cells in multipotent mesenchymal stromal cell cultures: current methods and future directions
Publication date: Available online 25 May 2019Source: CytotherapyAuthor(s): WEICHAO Zhai, DERRICK YONG, JEHAN JOMAA EL-JAWHARI, RICHARD CUTHBERT, DENNIS MCGONAGLE, MAY WIN NAING, ELENA JONESAbstractRegardless of their tissue of origin, multipotent mesenchymal stromal cells (MSCs) are commonly expanded in vitro for several population doublings to achieve a sufficient number of cells for therapy. Prolonged MSC expansion has been shown to result in phenotypical, morphological and gene expression changes in MSCs, which ultimately lead to the state of senescence. The presence of senescent cells in therapeutic MSC batches is und...
Source: Cytotherapy - May 26, 2019 Category: Cytology Source Type: research

Comparison of extraction methods and culture medium for umbilical cord lining- and wharton's jelly-derived mesenchymal stromal cells
ConclusionSix donated UC samples were disinfected, cut into discs and frozen in cryopreservant containing 10% DMSO. Samples were thawed at 37°C, then incubated in either media for 3 - 4 days at 37°C in 5% CO2.MSC were extracted from the incubated tissues and differentially treated to generate every permutation of cell source, extraction method and culture media (total conditions: 8). At every passage, cell doubling rate was calculated and morphology was captured. At Passage 6 (P6), trilineage differentiation and karyotype were evaluated by StemPro® differentiation kits and G-banding.MSC extracted from all condi...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Role Of Impeller Design For The Cultivation Of Stem Cells In Stirred-Tank Bioreactors
ConclusionWe have developed a novel 8-blade impeller to improve the cultivation of stem cells as aggregates and on microcarriers in the DASbox Mini Bioreactor System. Use of this impeller supported the formation and growth of cell aggregates in the bioreactor. Cell aggregate formation was more efficient than when using a conventional pitched-blade or Rushton-type impeller. Furthermore, microcarriers could be efficiently brought to and kept in suspension. Our results suggest that the impeller shape is an important parameter to consider when optimizing the agitation conditions in stirred-tank bioreactors. (Source: Cytotherapy)
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Bach impeller for suspension microcarrier cultures of mesenchymal stromal cells
ConclusionCulture of Wharton Jelly MSC at 1 litre with Solohill microcarriers showed 25% improved cell densities of the Bach impeller over the axial flow and paddle impellers: 4 × 10e5 cells/ml vs. 3 × 10e5 cells/ml. This was primarily due to extensive clumping of cell-microcarrier aggregates seen in the conventional impellers from day 3 onwards; whereas the laminar flow prevented aggregation up to the final day of harvesting.Significantly, when MSC were cultured on Cytodex microcarriers at 1 litre, cell yields increased dramatically to 15 × 10e5 cells/ml; viability and identity were equivalen...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

ADULT BOVINE PLATELET LYSATE-DERIVED SERUM “NeoSERA” IS SAFE, LESS ETHICAL AND POWERFUL ALTERNATIVE TO FETAL BOVINE SERUM FOR THE CULTURE OF MESENCHYMAL STEM CELLS
ConclusionNeoSERA is produced in Japan, considered to be a negligible bovine spongiform encephalopathy (BSE) risk country. We obtain adult donor bovine blood from animal less than 36 months old according to the regulation of EMA for BSE (EMA/410/01 rev.3). Using apheresis medical devices with closed disposable kits, sterile platelet-rich plasma (PRP) is collected from healthy donor bovine receiving a regular veterinary check. After stimulation and removal of coagulated fibrin by centrifugation, NeoSERA is collected in a completely closed system. To meet the scope of directives that apply to produce medicinal products from ...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Towards cell therapy of polycystic ovary syndrome (pcos): human mesenchymal stem cells secretome inhibits androgen production by pcos theca cells
Conclusion Material and MethodsTheca cells, collected from normal cycling and PCOS patient underwent ovariectomy, were seeded on culture dishes pre-coated with the extracellular matrix at a density of 6 × 104 cells/ml and cultured for 24 hours. Cells were then treated with either 3T3 conditioned media as a control (C) or Mesenchymal stem cells conditioned media (CM). The C and CM were prepared through collecting the serum-free supernatant of 3T3 and hMSCs at an optimal confluence, 48 hours later. The mRNA and protein expression of CYP17A1, CYP11A1, and DENND1A.V2 were quantified by RT-PCR and Western blot, while ...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Durable Lipopolysaccharide-Induced Potentiation of Mesenchymal Stem Cells
ConclusionIn the current study, we evaluated the feasibility of ex-vivo culture with LPS from E.coli O111:B4 as a mechanism of potentiating MSC secretion. Three independent human-derived bone marrow MSCs (BMSCs) were cultured ex-vivo in the presence of 5 µg/ml LPS. After 48 hours, conditioned medium (CM) was collected. The cells were washed and cultured for an additional 48 hours in complete medium. The CM was assayed for the presence of 42 different cytokines and growth factors using the multiplex bead platform.We found that compared to control cells not stimulated with LPS, generally ex-vivo culture with LPS signif...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Establishment of efficient differentiation conditions from tonsil-derived mesenchymal stem cells to motor neuron cells
ConclusionAfter the MN differentiating in our own methods, we confirmed that the expression of MN-related markers, including ISL1, HB9, ChAT in T-MSC-MNCs has increased compared to T-MSCs. Also, to test their functional efficiency, the change of acetylcholine secretion in the culture medium under the differentiation into T-MSC-MNCs was measured. Furthermore, by confirming the acetylcholine receptor clusters when co-culturing T-MSC-MNCs and human skeletal muscle cells, which was demonstrated the T-MSC-MNCs could form neuromuscular junctions. The functional improvements afforded by these T-MSC-MNCs are potentially useful in ...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Meeting the demands of msc production for clinical studies
ConclusionFor the trial, 3 lots of hCT-MSCs were manufactured from cord tissue obtained from 3 term healthy male babies delivered after elective routine C-section. After transporting the cord to the Robertson GMP Laboratory, the tissue was cut and digested in a tissue dissociator. The resulting cell suspension was plated in tissue culture flasks for P0 culture in Prime-XV MSC Expansion XSFM culture medium (Irvine Scientific) supplemented with 1% platelet lysate (PL). After harvest, P0 cells were cryopreserved, with a subset of cells expanded for P1 in XSFM without PL, and with a larger cryopreservation and expansion proces...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Comparative analysis of FBS containing media and serum free chemically defined media for adipose derived stem cells production
Conclusion MethodsMedia performance was compared with cell growth rate, accumulated cell number, population homogeneity and cell viability. Comparative analysis of adipose derived mesenchymal stem cells (ADSCs) was performed based on stem cell surface marker expression, differentiation potency, genetic stability, senescence analysis, mRNA expression change, and protein expression change.ResultsCulture using SFCDM provided more stable population doubling time (PDT), more homogenous cell population, and more cells in a shorter time compared to FBS containing media (FBSCM). The cultured ADSCs were shown to be positive for CD7...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Gamma irradiation of human platelet lysate: validation of efficacy for pathogen reduction and assessment of impacts on hpl performance
ConclusionIn this study, we validated the efficacy of gamma irradiation for virus inactivation. Four model viruses (BVDV, Reo3, HSV1, MMV) were chosen, per ICH/EMA guidelines, to represent a range of viruses with different genome, structure, size, and sensitivity to various chemical and physical agents. The virus spiked hPLs were gamma irradiated and the mean values of viral titers showed over 4 log10 reductions across all model viruses. The results demonstrated gamma irradiation is an effective viral reduction procedure for hPL.To assess the impacts of gamma irradiation on the long-term stability of hPL performance, we an...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Enhance hepatic differentiation of human Wharton's jelly–derived mesenchymal stromal cells by using sodium butyrate pre-treated
ConclusionHepatic differentiation of hWJ–MSCs was induced with 3–step differentiation protocol. The differentiated cells were examined for the expression of hepatic–specific markers and hepatocyte function by using immunocytochemistry, real–time reverse transcriptase–polymerase chain reaction (RT–PCR) and Periodic acid–Schiff (PAS) staining. Before hepatic differentiation of hWJ–MSCs, we identified embryonic definitive endodermal cells by using RT–PCR, which increase of CXCR4 mRNA level, after NaB pre–treated along with EGF and bFGF supplementation. Hepatic differ...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Safety Issues of Peribulbar Injection of Umbilical Cord Mesenchymal Stem Cell (UC-MSC) in Patients with Retinitis Pigmentosa
Conclusion MethodsThis study was conducted on 18 eyes of 18 volunteers with retinitis pigmentosa, The cells delivery with peribulbar injection method. Clinical examinitation, visus and fundus examination, multifocal electroretinography were performed before and after an peribulbar injection of approximately one-million UC-MSC. The patients were followed in 1 hour, 24 hours and 1 month for safety evaluation.Results: There are adverse events, like edema under the eyes, were observed in eyes all patients 1 hour after transplantation of UC-MSCs. The edema will disappear within 24 hours. These patients reported improvements in ...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Cartilage regeneration by autologous adipose-derived mesenchymal stem cells for the treatment of osteoarthritis
ConclusionHere we show that in twelve clinical cases, the cartilage was observed successfully regenerating and covering the defect region in knees after the intra-articular injection with adipose-derived mesenchymal stem cells (MSCs), and the treatment highly improves the symptoms of osteoarthritis. Basically, the adipose-derived MSCs from the patients are cultured and expanded to a specific amount with the optimized stem cell culture medium, the HELENE MEDIUM. After that, the cultured MSCs are directly injected into the injured knee joints of patients. The cartilage has gradually regenerated and been monitored by MRI phot...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Immunomodulation by mesenchymal stem cells is sex dependent
ConclusionHuman Passage 4 ADMSC (>30yrs old) from both male and female donors characterised via ISCT guidelines were assessed for in-vitro functionality and potency. Mitogen stimulated CD3+ T cells(>30yrs old) assessed via CFSE proliferation on FACS indicate a sexual dimorphism when donor sex ADMSC are co-cultured with same and opposite sex immune cells. ADMSC mediated T cell suppression indicates female ADMSC are more suppressive than their male counterparts(p=0.0002).Female ADMSC produce higher concentrations of pro-inflammatory cytokine mediated immunomodulatory markers like IDO1, IL-1RA and PGE-2 when pre-treated...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

The optimum temperatures and mediums storage for stability of umbilical cord derived mesenchymal stem cell (uc-msc) and stromal vascular fraction (svf) viability
ConclusionBoth of UC-MSCs and SVF were divided into 4 groups with two variables temperature and medium storage. Cells have been evaluated in room temperature (RT) (18-22°C) and cold temperature (2-8°C) with 0.9% NaCl and conditioned medium (UC-MSCs) or autologous serum (SVF). The viability from each group was assessed day by day using Tryphan Blue.At room temperature, viability of cells in both storage mediums ware significantly decreased while stored at cold temperature the viability was more stable. The highest viability of UC-MSCs at RT was obtained from cells in 0.9% NaCl solution (77% viable). However, the hig...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

A novel serum- and xeno-free medium for the expansion of human mesenchymal stromal/stem cells.
ConclusionHere, we show the performance of a novel serum- and xeno-free cell culture medium developed for the expansion of hMSCs. This medium enables scale-up from planar culture to suspension culture in stirred tank bioreactors using microcarriers. In addition to consistent and good growth performance in both planar and suspension systems, the hMSCs expanded in this culture medium maintain all the characteristics of multipotency. The medium is manufactured under cGMP conditions using raw materials selected for high quality and suitability for manufacturing applications and does not contain phenol red.This novel serum- and...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Evaluation of human platelet lysate (hpl) as a substitute for foetal bovine serum (fbs) and recombinant proteins for the growth and maintenance of bone marrow derived mesenchymal stromal cells (bm-msc)
ConclusionThe growth and phenotype of BM-MSC cultured in either apheresis HPL or commercial HPL were comparable, suggesting apheresis HPL may prove a suitable alternative to FBS. A simpler preparation protocol of HPL for the end users will increase the likelihood of introducing HPL into current laboratory practice. The potential of pooled HPL warrants further investigation. (Source: Cytotherapy)
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Dissecting the molecular pathways of apoptosis in mesenchymal stromal cell therapy
ConclusionUsing pharmacological compounds that selectively inhibit pro-survival proteins to induce apoptosis in MSCs, we show that mouse and human bone marrow (BM) MSCs engage different molecules for survival. In addition, we are genetically manipulating the apoptotic pathways in mouse and human BM MSCs to gain a greater understanding of how these cells exert their therapeutic effects in vivo, an outstanding question in MSC therapy. (Source: Cytotherapy)
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Rapid expansion of Mesenchymal Stem/Stromal Cells using optimized media supplemented with human platelet lysate PLTMax® or PLTGold®, suitable for cGMP expansion at large scale.
ConclusionIn traditional monolayer expansion systems, we have demonstrated growth of adipose derived and bone marrow derived MSCs using an optimized, fully cGMP compliant culture system. We have found that MSC NutriStem® XF Basal Medium (Biological Industries USA, Inc, Cromwell, CT) supplemented with PLTMax® or PLTGold® exceeds the performance of other conventional media products, obtaining up to 5 × 108 cells in only 5 passages (5 times more cells than with other commercially available products). Furthermore, when we use MSC NutriStem® XF Complete Medium supplemented with PLTMax® or PLTGold&r...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Tide motion bioreactors for large-scale cultivation and expansion of human mesenchymal stem cells
ConclusionEsco Aster has leveraged on the Tide Motion bioreactors to establish a robust and scalable platform using macrocarriers to meet the demands for future clinical therapies. MSCs isolated from different lineages were seeded and allowed to expand within PET macrocarriers while cell counts were measured and monitored daily. Extracellular matrices secreted by MSCs was observed in culture, which promote more physiological-like conditions and aid in the expansion of MSCs and maintaining their physiological characteristics. Throughout culture periods, cell culture conditions were monitored, with bioprocess parameters such...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Stability Characteristic of Cryopreserved Human Umbilical Cord Wharton's Jelly–Derived Mesenchymal Stromal Cells
ConclusionWJ–MSCs were isolated at a 100% successful rate and characterized by flow cytometry for CD34, CD45, CD73, CD90, CD105, CD106 and STRO–1. Growth kinetics characteristics were investigated by population doublings (PD), clinical large–scale expansion and high viability maintenance. MSCs were assessed for differentiation potential to cartilage, adipose and bone. Flow cytometry analysis showed no differences in expression of CD34, CD45, CD73, CD90, CD105, CD106 and STRO–1 among all cryopreserved WJ–MSCs for six years (2013–2018: n=30). All years of cryopreserved WJ–MSCs mainta...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Comparison of Viability and Proliferation Potency of Stromal Vascular Fraction Stored at Different Temperature
ConclusionA total of 70 ml donor-adipose tissue was divided into two different groups, room temperature (RT) and 4-8°C. SVF was isolated using manual method. Each group was process after stored for 4, 5, 6, 7, and 12 days. After it passed each incubation time, adipose samples were processed enzymatically until SVF was obtained. The viability of SVF stored at RT was decreasing until 40%, but relatively constant when stored at 4-8°C (>75%). Nevertheless, the sample stored at RT revealed to have better proliferation and colonization than sample stored at 4-8°C. The sample retained the proliferation capability w...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Stromal vascular fraction (svf) therapy for treatment of various diseases: delivering safety of the first patented svf technique in indonesia
ConclusionStromal vascular fraction (SVF) therapy has been performed over the past seven years to treat 493 patients by our group in five clinical centers. SVF was isolated from the lipoaspirate collected from each respective patient through a simple manual liposuction procedure. For each patients, blood was drawn as well to isolate platelet-rich plasma (PRP). SVF and PRP were administered to all subjects by intravenous (IV) injection in an autologous manner, with minority group within the population received an additional spinal, intra-articular, or subcutaneous injections, or combination of several injection sites. The t...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Brain-death donors as an alternative source of human stromal mesenchymal cells for cell-based therapy
ConclusionAdequate procedures were developed for BM collection and processing, as well as for in vitro culture, immunophenotyping and differentiation. Fourteen donor-matched BM samples (IC and F) were processed from seven deceased donors (Table 1). Our results show that it is possible to obtain viable hMSC from both sites. Although the total number of mononuclear cells (MNC) per gram of BM appears to be greater in the CI than in F, no statistically significant differences were observed between both sources. Neither the total content of fibroblastoid colony-forming units (CFU-F) per gram of BM nor the amount of CFU-F per MN...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Challenges in developing an off-the-shelf cell therapy for ACLF and NASH
ConclusionThe dosing for this indication is likely to be in the order of 100 million cells per infusion. Large numbers of cells thus need to be manufactured and using standard cell culture techniques would make it too expensive and labour intensive. A standard expansion protocol in flasks or cell factories is an open process, and therefore run in expensive clean rooms to avoid the risk of contamination. We have explored several state-of-the art-bioreactor systems, and various types of microcarriers in stirred tanks from different manufacturers. We have tested these systems to optimize the complete workflow of HepaStem cult...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Will not be presented
Publication date: May 2019Source: Cytotherapy, Volume 21, Issue 5, SupplementAuthor(s): (Source: Cytotherapy)
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Effect of heparin on proliferation mesencymal stem cell
ConclusionIn this study, the addition of heparin to mesenchymal stem cell culture medium with concentration of each treatment group 0 IU / ml (as control); 0.30 IU / ml; 0.61 IU / ml; 1.25 IU / ml; 2.5 IU / ml; 5 IU / ml; and 10 IU / ml. The culture was carried out for 15 days. On the 1st day; the 3rd; 7th; 10th; and 15th, do MTT test.In the study, the highest live cell count was in the heparin treatment group of 0.61 IU / mL (1.9 × 103 cells / mL) p = 0.000.ConclusionAdding heparin in culture medium, increased the proliferation of mesenchymal stem cells with optimum concentrations of 0.61 IU / mL on day 7th ...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

A Safe and Simple Approach to Obtaining Sufficient Cytokine Induced Killer (CIK) Cells from Critically-ill Cancer Patients Without the Need for Apheresis or Peripheral Mobilization
Conclusion Methodology60ml of peripheral venous blood was withdrawn from 9 patients (M=5, F=4) with mean age of 62±11 years old and 1 healthy donor (male, 45 years) using simple venipuncture technique. The samples were isolated and induced using interferon-gamma (IFN-γ), anti-CD3 antibody, recombinant human interleukin-2 (IL-2) for up to 21 days.ResultsWe successfully isolated and expanded CIK from all 10 blood samples. The number of CIK was significantly increased from Day 1 to Day 21 of cell expansion (0.12±0.08 × 109 vs. 7.46±1.85 × 109 cells, p
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Arthroscopic Evidence of Cartilage Regeneration in Severe Knee Cartilage Defects and Osteoarthritis Following Treatment with Allogeneic Umbilical Cord-Derived Mesenchymal Stromal Cells (Chondrocell-Ex)
Conclusion MethodThis is a single-arm, open-label, compassionate study at the Universiti Kebangsaan Malaysia Medical Centre (UKKMC) with 12 months follow-up period. Subjects with knee articular cartilage defect ICRS 3 diagnosed via diagnostic arthroscopy were treated with allogeneic UC-MSCs (Chondrocell-Ex, Cytopeutics, Malaysia) and assessed in five subsequent visits. During the visits, assessment was done following Visual Analog Score (VAS), International Knee Documentation Committee (IKDC), Tegner Activity Score (TAS), and Knee injury and Osteoarthritis Outcome Score (KOOS) scoring. At the end of the study, subjects und...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Mesenchymal Stromal Cells (Cyto-MSC) are Well-tolerated and Associated with Renal Function Improvement in Patients with Chronic Kidney Disease: An Observational Retrospective Study
Conclusion MethodologyThere were 4 clinical trials approved at the National University Malaysia in collaboration with Cytopeutics (Cyberjaya, Malaysia) which provided the mesenchymal stromal cells (Cyto-MSC) for treatment of acute stroke, heart failure, diabetes with critical limb ischemia and severe deforming osteoarthritis. All patients received 65-130 × 106 Cyto-MSC intravenously. Severity of chronic kidney disease is based on baseline estimated glomerular filtration rate (eGFR) and serum creatinine which are markers of renal dysfunction. These and other blood tests including fasting blood sugar (FBS) and Hba1...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Dose-dependent Immunomodulatory Effect of Intravenous Allogeneic Umbilical Cord-Derived Mesenchymal Stromal Cells (Cytopeutics CLV-100) Infusion in Healthy Volunteers
Conclusion MethodologyUmbilical cord samples were collected after delivery of full-term, healthy babies with written consent from both parents. All 3 generations (newborn, parents and grandparents) were screened for genetic mutations, infections, cancers and other inherited diseases. Samples were transferred to a certified Good Manufacturing Practice laboratory for processing. Subjects were infused with either low dose (LD, 65 million cells) or high dose (HD, 130 million cells) of CLV-100 followed up for 6 months. We measured cytokines level using ELISA including the acute phase reactant high-sensitivity C-reactive protein...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Cytopeutics® Umbilical Cord Mesenchymal Stromal Cells (Cyto-MSC) for Patients with Grade II–IV Acute Graft-Versus-Host Disease: A Phase I/II Clinical Study – Protocol Overview
Conclusion Study DesignThis is a phase I/II clinical study involving patients who underwent an allogeneic HSCT for malignant or non malignant haematological disorders and developed grade II-IV acute GVHD. A total of 40 eligible patients will be recruited in this study.For Phase I open labelled study, 5 eligible patients will be recruited to receive Cyto-MSC (5 × 106 MSC per kg bodyweight) and standard treatment. Meanwhile, for Phase II double blinded placebo controlled study, another 35 eligible patients will be recruited and randomized into 2 study groups where 15 patients will be assigned into Group A to receiv...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Standardized qc assays and large scale expansion of pluripotent stem cells using an automated closed system.
ConclusionHere, we established a workflow encompassing stable expansion of hPSCs using a xeno-free cultivation medium, assessment of pluripotency using a defined marker combination and assessment of differentiation potential based on lineage-specific, complete media both combined with quantitative, multicolor flow cytometry analysis to characterize and quality control the cultivated hPSCs, as well as cryopreservation of hPSCs using an animal component-free, chemically defined cryopreservation media.Following this workflow, hPSCs could be stably expanded over 20 passages with persistent, high expression of pluripotency mark...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Efficient GMP compliant expansion of mesenchymal stromal cells (MSCs) from umbilical cord, bone marrow and adipose tissue using a closed cultivation system.
ConclusionBM-MSCs were isolated via DGC and subsequent adherence to plastic using semi-automatic feeding, harvesting and reseeding procedures.AT-MSCs as well as UC-MSCs were isolated and expanded from single-cell suspensions obtained after enzymatic digestion. UC was digested using an optimized and automated procedure using the gentle MACSTM Dissociator. Subsequently, MSCs were isolated from single cell suspension using the flexible cultivation procedure of the CliniMACS Prodigy® Adherent Cell Culture System.Resulting cells displayed an MSC specific phenotype as defined by the ISCT consortium. Furthermore MSCs revealed...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Identification and Characterization of iPS Clones via Automated Live-Imaging and In-Process Analysis for Quality Control using High Throughput Robotic System
ConclusionWe used DF6-9-9T.B hiPSC cell line, reprogrammed skin fibroblasts and peripheral blood mononuclear cells to develop our image processing parameters and picking and weeding protocols. To define and quantify critical quality attributes (CQAs) of “completely” reprogrammed iPS cells in systematic repeatable and reproducible manner, the Cell XTMPlatform is integrated with ColonyzeTM4.0, a very user-friendly WindowsTM-based automated quantitative cell and colony analysis software. The principles and nomenclature enabling this approach are outlined in ASTM Standard F2944-12. To automate manipulation of iPS c...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Characterization of Human Cartilage-Derived Progenitor Heterogeneity for Cellular Therapy Strategies Using Time-Lapse Phase-contrast Imaging
ConclusionArticular cartilage (Outerbridge grade 1-2) obtained from nine knee arthroplasty patient's were enzymatically digested to isolate cells for 2-D cell culture assay. Large field of view phase contrast images were acquired hourly to capture images of the progenitors and their progeny (colony) for standardized ASTM-based automated colony-forming unit (CFU) analysis to define and quantify CQAs of the stem/progenitor cells and their progeny, which include: 1) lag-time for first division, 2) circularity, 3) area, 4) feret's diameter, 5) effective proliferation rate, and 6) colony density.Of the 225 colonies obtained fro...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

High Throughput Performance-Based Selection and Expansion of Clonal Progenitor Populations Resident in Human Articular Cartilage to Characterize Sub-populations for Effective Cartilage Stem Cell Therapies
ConclusionArticular cartilage (Outerbridge grade 1-2) obtained from six knee arthroplasty patient's were enzymatically digested to isolate cells for 2-D cell culture assay. Large field of view images were acquired daily to capture images of the progenitors and their progeny for standardized ASTM-based automated image analysis to define and quantify CQAs of the stem/progenitor cells. Based on the CQAs identified, 24 clonal populations from each patient were picked using Cell XTMrobotic device in rapid, precise, repeatable and rigorously documented manner. Of the 24 clonal colonies, twelve fastest growing clones were expande...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Inicial Request Agreement with Researchers of ANMAT, (Argentinean FDA), to Present Clinical Trials to Obtain Approval and Registers of Complex Manipulated Cellular Therapy Products and Products to be used in Tissue Engineering
Conclusion MethodsThe main obstacle for clinical trial regulators was the lack of information of how to homologate the knowledge of Drug Characterization, Pharmacokinetics, Pharmacodynamics and Safety the analogic study for cellular products.To approach this problem Center for Research on Tissue Engineering and Cellular Therapy (CIITT) presented a full documentation for a clinical trial. It was written following FDA recommendations and theoretical concepts, presented during the Global Regulatory Perspective (GRP) annual conferences.Then, a careful discussion with the regulators was sustained, satisfying all the concerns th...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Specific Effector Cell (EC) Immunomodulation improves the repair function of adipose Mesenchymal Stroma Cells (aMSC) on an experimental model of Grade3-4 Osteoarthrosis (OA). Comparation with other current therapies
Conclusion Methods44Wistar rats (250 g)were intra articular (IA) injected with a0.2% solution of mono iodine acetate (MIA, MERK Lab). After 7 days, 4 animals were euthanized and injected joins, as well its contralateral, were collected for 0-day histology controls. The rest of animals were divided into 5 groups of 8 animals each. Group 1 was kept w/o treatment as a Control Group,Group 2 IA treated with syngeneic aMSC, Group 3 with syngeneic activated EC, Group 4 with a co-culture of EC+aMSC and Group 5 with syngeneic PRP. 4 animals of each group were euthanized at day 7, and the remain 4 rats were euthanized for day 14. aM...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Reducing the dependency of operator-based counting methods during the manufacturing process of cell and gene therapies.
ConclusionA series of dilution was performed to assess T-cell and mesenchymal stem/stromal cell (MSC) concentration accuracy and precision measured using one manual method (haemocytometer) and two automated methods. The linearity of each counting method in regards to cell concentration was also compared. Additionally, viability and diameter accuracy together with precision across the three different counting systems were also assessed.Regarding cell concentration accuracy, the automated method I was shown to be comparable to the haemocytometer. The reads obtained from the automated method II indicated superior cell concent...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Will not be presented
Publication date: May 2019Source: Cytotherapy, Volume 21, Issue 5, SupplementAuthor(s): (Source: Cytotherapy)
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research

Development of an abbreviated protocol for the polarization and characterization of therapeutic inflammation-resolving monocytes/macrophages
Publication date: May 2019Source: Cytotherapy, Volume 21, Issue 5, SupplementAuthor(s): M. Chan, A. Gomez-Aristizabal, R. Rabani, R. Gandhi, W. Marshall, N. Mahomed, S. ViswanathanBackground & AimMonocytes/macrophages (MΦs) are innate immune cells that exist on a spectrum of pro-inflammatory to inflammation-resolving phenotypes. Their polarization is mediated in vivo through the local microenvironment or ex vivo through exogenous factors. Such ex vivo polarized cells have been used in early clinical trials for a variety of immunosuppressive or reparative indications (e.g. spinal cord injury, liver cirrhosis, transp...
Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research