A Defined and Xeno-Free Culture Method Enabling the Establishment of Clinical-Grade Human Embryonic, Induced Pluripotent and Adipose Stem Cells

作者：Kristiina Rajala, Bettina Lindroos, Samer M. I. Hussein, Riikka S. Lappalainen, Mari Pekkanen-Mattila, José Inzunza, Björn Rozell, Susanna Miettinen, Susanna Narkilahti, Erja Kerkelä, Katriina Aalto‐Setälä, Timo Otonkoski, Riitta Suuronen, Outi Hovatta, Heli Skottman · 发表于：PLoS ONE · 年份：2010 · DOI：10.1371/journal.pone.0010246 · 被引用次数：166 · 研究领域：Pluripotent Stem Cells Research、Tissue Engineering and Regenerative Medicine、Mesenchymal stem cell research

BACKGROUND: The growth of stem cells in in vitro conditions requires optimal balance between signals mediating cell survival, proliferation, and self-renewal. For clinical application of stem cells, the use of completely defined conditions and elimination of all animal-derived materials from the establishment, culture, and differentiation processes is desirable. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report the development of a fully defined xeno-free medium (RegES), capable of supporting the expansion of human embryonic stem cells (hESC), induced pluripotent stem cells (iPSC) and adipose stem cells (ASC). We describe the use of the xeno-free medium in the derivation and long-term (>80 passages) culture of three pluripotent karyotypically normal hESC lines: Regea 06/015, Regea 07/046, and Regea 08/013. Cardiomyocytes and neural cells differentiated from these cells exhibit features characteristic to these cell types. The same formulation of the xeno-free medium is capable of supporting the undifferentiated growth of iPSCs on human feeder cells. The characteristics of the pluripotent hESC and iPSC lines are comparable to lines derived and cultured in standard undefined culture conditions. In the culture of ASCs, the xeno-free medium provided significantly higher proliferation rates than ASCs cultured in medium containing allogeneic human serum (HS), while maintaining the differentiation potential and characteristic surface marker expression profile of ASCs, although signific...