Abstract
Culture methods and environmental cues of human pluripotent stem cells
Highlights
Standard human pluripotent stem cells cultures have supportive cells such as inactivated mouse embryonic fibroblast (MEF) feeder cells that aid cell growth, secrete several important growth factors into the medium, which help maintain pluripotency and prevent differentiation [1]
Feeder based cultures are suitable for routine maintenance of human pluripotent stem cells (hPSCs) colonies, and MEFs are the most frequently used feeder cells, because they support the robust growth of all types of embryonic stem cells as colonies [2]
Since MEFs have complex and undefined heterogeneity, a variety of human cell types such as human fibroblasts, tubal, foreskin, and bone marrowderived stromal cells can be used as feeder cells instead of MEF [2]
Summary
Standard human pluripotent stem cells (hPSCs) cultures have supportive cells such as inactivated mouse embryonic fibroblast (MEF) feeder cells that aid cell growth, secrete several important growth factors into the medium, which help maintain pluripotency and prevent differentiation [1]. HPSCs can be cultured on NCM at the surface of human recombinant laminin-512 coated polystyrene without the use of a ROCK inhibitor, and generally hPSCs under these growth conditions have genetic stability and pluripotency [3]. The advantages of this culture are the feeder-free, adjustable growth rate, the production of homogeneous hPSCs, rapid cell growth, rapid (2-4 days) cell recovery after cryopreservation (vs 1 to 3 weeks of colony culture). HPSCs grown with NCM are highly effective in teratoma formation and can be converted into colony-type culture if grown as clumps [2]
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