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 expanded to 20 doublings for trilineage differentiation assay and RNA sequencing analysis.Preliminary assessment indicated wide variation in morphological and biological characteristics of progenitor and their progeny: circularity (median: 0.665; range:0.15-0.93), area (median: 116.4µm2; range:51.9-204.2µm2), doubling time (median:32.9h; rangse:26.9-41.2h) and colony density (median:6.5%;range:2.3-20.7%). Differences seen between two representative clonal populations with respect to the trilineage differentiation potential and gene expression seen at 20 doublings is shown in Figure 1 and 2 respectively. A complete detailed assessment of all the clonal populations and their culture-expanded population (n=72) will help us identify the CQAs that will aid in selection of quality clones.An improved understanding of the heterogeneity of progenitor cells resident in adu...
Source: Cytotherapy - Category: Cytology Source Type: research