Establishment of an automated patch-clamp platform for electrophysiological and pharmacological evaluation of hiPSC-CMs

Publication date: Available online 18 November 2019Source: Stem Cell ResearchAuthor(s): Wener Li, Xiaojing Luo, Ying Ulbricht, Michael Wagner, Christopher Piorkowski, Ali El-Armouche, Kaomei GuanAbstractHuman induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) have evolved into widely used and reliable cell sources for modeling cardiovascular channelopathies and for drug safety pharmacology. However, the electrophysiological and pharmacological applications of hiPSC-CMs are hampered by manual patch-clamp technique, which is labor-intensive and generates a low-output. The automated patch-clamp technique is showing potential to overcome this problem. Here, we describe a new dissociation method, with which we can harvest a vast number of single relaxed hiPSC-CMs with smooth membrane suited for automated patch-clamp. Using the automated whole-cell patch-clamp technology, we report a high success rate for cell capture and whole-cell access (around 70%). We are able to identify and record several currents and paced action potentials (APs) with different success rates, including Na+ current (INa), L-type Ca2+ current (ICaL), two specific K+ currents, the transient outward K+ current (Ito) and the inward rectifier K+ current (IK1). Moreover, we successfully applied dynamic current-clamp to virtually increase IK1 for AP recordings. Our study suggests that automated patch-clamp technology could be used to investigate the relevant ionic currents and APs in hiPSC-CMs. The co...
Source: Stem Cell Research - Category: Stem Cells Source Type: research

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In this study, the enhanced mice live somewhat longer than their unmodified peers, though not as much longer as is the case for the application of telomerase gene therapy. The mice do also exhibit reduced cancer risk, however. The scientists here class telomere shortening as a cause of aging, which is not a point universally agreed upon. Reductions in average telomere length in tissues looks much more like a downstream consequence of reduced stem cell activity than an independent mechanism. Researchers obtain the first mice born with hyper-long telomeres and show that it is possible to extend life without any geneti...
Source: Fight Aging! - Category: Research Authors: Tags: Newsletters Source Type: blogs
This study aims to explore whether mesenchymal stem cells (MSCs) affect RDS in newborn swines via the Janus kinase-signal transducers and activators of transcription (JAK-STAT) signaling pathway by the establishment of the model of the disease. MATERIALS AND METHODS: The phosphorylation of the JAK-STAT signal transduction proteins was first detected via Western blotting to verify the regulatory effect of MSCs on RDS in newborn swines through the JAK-STAT signaling pathway. Then, the Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was utilized to analyze the influences of the injection of MSCs into the blood of...
Source: European Review for Medical and Pharmacological Sciences - Category: Drugs & Pharmacology Tags: Eur Rev Med Pharmacol Sci Source Type: research
In this study, we investigated the molecular and antigenic properties of T. parva antigen Tp9 expressed by mammalian cells. Data indicate that Tp9 contains a signal peptide that is weakly functional in mammalian cells. Thus, Tp9 secretion from mammalian cells increased 10-fold after the native signal peptide was replaced with the human tissue plasminogen activator signal peptide (tPA). Sera from all T. parva-immune cattle recognized this recombinant, secreted Tp9. Additionally, PBMC from ITM-immunized cattle produced significant (p 40.2°C) were treated with flunixin meglumine, and those that developed dyspnea were trea...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Geng-Hong Xia1†, Chao You1,2†, Xu-Xuan Gao1, Xiu-Li Zeng1, Jia-Jia Zhu1, Kai-Yu Xu3, Chu-Hong Tan1, Ruo-Ting Xu1, Qi-Heng Wu1, Hong-Wei Zhou3, Yan He4*‡ and Jia Yin1*‡ 1Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou, China 2Department of Neurology, The First People's Hospital of Zunyi, Zunyi, China 3State Key Laboratory of Organ Failure Research, Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China 4Microbiome Medicine Center, Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical Univ...
Source: Frontiers in Neurology - Category: Neurology Source Type: research
In this study, we address these issues, as well as the technical limitations of previous studies, by using RNA-Seq analysis to compare the transcriptomes of freshly isolated murine LN-associated LEC and BEC (LN-BEC) as well as freshly isolated LEC from the diaphragm (D-LEC) as representative of peripheral tissue lymphatics. RNA-Seq has greatly improved the analysis of whole transcriptomes with higher sensitivity and dynamic range coupled to lower technical variations compared to microarrays and quantitative PCR (19, 20). Our work provides an important resource for further exploration of endothelial cell functionality in di...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Conclusions Altogether, this review summarized the current knowledge of the polarization of macrophages and their regulatory mechanisms at different stages of pregnancy, as well as the roles of these cells in pathological processes. Although current evidence provides a compelling argument that macrophages are important in pregnancy, our understanding of the roles and mechanisms of macrophages in pregnancy is still rudimentary. Since macrophages exhibit functional plasticity, they may be ideal targets for therapeutic manipulation during pathological pregnancy. Additional studies are needed to better define the functions an...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
This study also identified that the depletion of Langerin+CD8+ cDC1 resulted in diminished protective immune responses against intravenous Mycobacterium bovis infection. Langerin+CD8+ cDC1-depleted mice displayed increased bacterial loads, due to decreased IL-12 production in combination with delayed and diminished CD8+ T cell responses (77). Interestingly, although CD8+ T cell responses recovered over time, the bacterial load continued to increase and could not be controlled. This indicates that early immune priming effects by Langerin+CD8+ cDC1 are essential for the fate of the immune response (77), which was also found ...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
This study was supported in part by the National Cancer Center Research and Development Fund (25-A-7 and 28-A-8); Health and Labor Science Research Grants for Clinical Research, Japan; and joint research funding from Takeda Pharmaceutical Co, Ltd.; Noile-Immune Biotech Inc.; Ono Pharmaceutical Co., Ltd.; BrightPath Biotherapeutics Co., Ltd.; and Sysmex Co., Ltd. This study was performed as part of a research program of the Project for Development of Innovative Research on Cancer Therapeutics (P-Direct), Ministry of Education, Culture, Sports, Science and Technology of Japan. Conflict of Interest Statement TN, TS, and TY ...
Source: Frontiers in Oncology - Category: Cancer & Oncology Source Type: research
Purpose of review Human kidney development and the mechanisms of many kidney diseases are incompletely understood partly due to the lack of appropriate models. Kidney organoids derived from human pluripotent stem cells (hPSCs) are a new and rapidly developing in-vitro system covering the window of early nephrogenesis and having the capacity for disease modelling. The application of global analytic tools such as RNA sequencing and proteomics is providing new and unexpected insights into kidney organoids with relevance for development and disease. In this review, we focus on the most significant advances in the field over ...
Source: Current Opinion in Nephrology and Hypertension - Category: Urology & Nephrology Tags: RENAL IMMUNOLOGY AND PATHOLOGY: Edited by Agnes B. Fogo Source Type: research
Authors: Li L, Sun Y, Zhang N, Qiu X, Wang L, Luo Q Abstract Sufentanil is a type of opioid analgesic and is usually used to facilitate painless labor in combination with the local anesthetic ropivacaine. One aim of the current study was to investigate the effects of sufentanil and ropivacaine on umbilical cord mesenchymal stem cells (UCMSCs). A second aim of this study was to determine whether sufentanil attenuated the cytotoxicity of ropivacaine in vitro. UCMSCs were divided into 3 groups: one was treated with ropivacaine at a concentration of 50, 100, 200, or 400 μg/mL, another was treated with sufentanil at ...
Source: BioScience Trends - Category: Biomedical Science Tags: Biosci Trends Source Type: research
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