Frequently Asked Questions on the Maintenance of hPSCs



Reagent Considerations

How do I choose the optimal TeSR™ media for my research applications?

Can I transition cells cultured in other feeder-free media to mTeSR™1?

Reagent Preparation

Why should TeSR™ supplement be thawed at room temperature or in the refrigerator instead of at 37°C?

My TeSR™ medium supplement appears slightly turbid after thawing. Can I still use it?

Is it necessary to add antibiotics to the media?

Why is it necessary to use non-tissue culture-treated cultureware for Vitronectin XF™(Catalog #07180)?

Passaging and Maintenance

Which is better, non-enzymatic dissociation or enzymatic dissociation?

Should I passage my hPSCs as aggregates or as single cells?

How does one determine the optimal number of aggregates to plate?

When is ROCK inhibitor required?

When is spontaneous differentiation a problem?



Related Resources

References

  1. Ludwig TE et al. (2006) Feeder-independent culture of human embryonic stem cells. Nat Methods 3(8): 637–46.
  2. Ludwig TE et al. (2006) Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol 24(2): 185–7.
  3. Chen G et al. (2011) Chemically defined conditions for human iPSC derivation and culture. Nat Methods 8(5): 424–9.
  4. Draper JS et al. (2004) Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol 22(1): 53–4.
  5. Buzzard JJ et al. (2004) Karyotype of human ES cells during extended culture. Nat Biotechnol 22(4):381–2.
  6. Watanabe et al. (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25(6): 681-6.
  7. Beers et al. (2012) Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions. Nat Protoc 7(11): 2029–40.
  8. Närvä et al. (2017) A Strong Contractile Actin Fence and Large Adhesions Direct Human Pluripotent Colony Morphology and Adhesion. Stem Cell Reports 9 (1): 67-76.

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