A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells

• Published in: PloS one Vol. 11; no. 9; p. e0161229
• Main Authors: Ahmadian Baghbaderani, Behnam, Tian, Xinghui, Scotty Cadet, Jean, Shah, Kevan, Walde, Amy, Tran, Huan, Kovarcik, Don Paul, Clarke, Diana, Fellner, Thomas
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 08.09.2016; Public Library of Science (PLoS)
• Subjects: Animals; Anticoagulants; Biology and Life Sciences; Biomarkers - metabolism; Cell culture; Cell Culture Techniques - methods; Cell Proliferation - drug effects; Cell Shape - drug effects; Cells, Cultured; Cultivation; Culture Media - pharmacology; Cyclic GMP; Cytokines; Ectoderm; Embryos; Endoderm; Fibroblast Growth Factor 2 - pharmacology; Fibroblasts; Growth factors; Humans; Manufacturing; Medicine and Health Sciences; Mesoderm; Methods; Mice, SCID; Plasmids; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - drug effects; Pluripotent Stem Cells - metabolism; Propagation; Regenerative medicine; Stem cell research; Stem cells; Time Factors; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0161229

Human pluripotent stem cells (hPSCs) present an unprecedented opportunity to advance human health by offering an alternative and renewable cell resource for cellular therapeutics and regenerative medicine. The present demand for high quality hPSCs for use in both research and clinical studies underscores the need to develop technologies that will simplify the cultivation process and control variability. Here we describe the development of a robust, defined and xeno-free hPSC medium that supports reliable propagation of hPSCs and generation of human induced pluripotent stem cells (hiPSCs) from multiple somatic cell types; long-term serial subculturing of hPSCs with every-other-day (EOD) medium replacement; and banking fully characterized hPSCs. The hPSCs cultured in this medium for over 40 passages are genetically stable, retain high expression levels of the pluripotency markers TRA-1-60, TRA-1-81, Oct-3/4 and SSEA-4, and readily differentiate into ectoderm, mesoderm and endoderm. Importantly, the medium plays an integral role in establishing a cGMP-compliant process for the manufacturing of hiPSCs that can be used for generation of clinically relevant cell types for cell replacement therapy applications.