AI Article Synopsis
- Enzymes traditionally used for passaging human pluripotent stem cells can harm the cells by digesting surface proteins, and alternative methods that lack divalent cations may cause cell death.
- The study finds that magnesium (Mg²⁺) and calcium (Ca²⁺) ions can effectively manage cell adhesion and detachment in hPSCs without using enzymes, allowing for healthier cell handling.
- This new enzyme-free method preserves more cell integrity and facilitates the collection of undifferentiated cells in larger clumps, making the culture process safer and easier.
Article Abstract
Enzymes used for passaging human pluripotent stem cells (hPSCs) digest cell surface proteins, resulting in cell damage. Moreover, cell dissociation using divalent cation-free solutions causes apoptosis. Here we report that Mg(2+) and Ca(2+) control cell-fibronectin and cell-cell binding of hPSCs, respectively, under feeder- and serum-free culture conditions without enzyme. The hPSCs were detached from fibronectin-, vitronectin- or laminin-coated dishes in low concentrations of Mg(2+) and remained as large colonies in high concentrations of Ca(2+). Using enzyme-free solutions containing Ca(2+) without Mg(2+), we successfully passaged hPSCs as large cell clumps that showed less damage than cells passaged using a divalent cation-free solution or dispase. Under the same conditions, the undifferentiated and early-differentiated cells could also be harvested as a cell sheet without being split off. Our enzyme-free passage of hPSCs under a serum- and feeder-free culture condition reduces cell damage and facilitates easier and safer cultures of hPSCs.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3983606 | PMC |
| http://dx.doi.org/10.1038/srep04646 | DOI Listing |
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