Journal: International Journal of Molecular Sciences
Article Title: End-to-End Platform for Human Pluripotent Stem Cell Manufacturing
doi: 10.3390/ijms21010089
Figure Lengend Snippet: Human iPSCs expanded in a bioreactor have typical hiPSC morphology and express hPSC-associated markers post-harvest. ( A ) Phase contrast images of iPSCs expanded in a bioreactor plated either as single cells after cell release from MCs (left panel), or as clusters of cells on MCs (right panel). Magnification, 100×; Scale bar: 100 µm; ( B ) Immunofluorescence staining of OCT3/4, NANOG, SSEA-4, TRA-1-60 and TRA-1-81 of iPSCs expanded in a bioreactor and plated onto 2D cell culture plates. Representative images from two different cell lines are shown. Magnification, 100×; ( C ) Quantitative analysis of hPSC-associated markers OCT3/4, SSEA4, TRA-1-60 and TRA-1-81 by flow cytometry of RTiPSC3B and LiPSC18R cells expanded in a bioreactor. Data obtained from either three independent runs (RTiPSC3B) or two independent runs (LiPSC18R). Data plotted are mean ± standard deviation (SD); ( D ) Pluripotency of RTiPSC3B cells expanded in a bioreactor demonstrated by embryoid body (EB) formation followed by immunofluorescence staining for germ layer-specific markers. Magnification, 100× (ectoderm, endoderm), 200× (mesoderm). ( E ) Representative immunofluorescence staining for lineage-specific markers RTiPSC3B and LiPSC18R cell lines. Magnification, 100× (Neural stem cells, definitive endoderm), 200× (cardiomyocytes)
Article Snippet: After nucleofection, the cells were plated onto 6-well plates pre-coated with L7TM hPSC Matrix, in priming medium containing 0.5 mM sodium butyrate (ReproCELL USA, Inc., 04-0005, Beltsville, MD, USA) to enhance reprogramming efficiency.
Techniques: Immunofluorescence, Staining, Cell Culture, Flow Cytometry, Standard Deviation