Table 1. Passaging protocols for hPSC culture.
| Passage Protocols | Culture Conditions | References | ||||
|---|---|---|---|---|---|---|
| Divalent cations | Enzymes | Manipulation | Cell clump size | Serum replacement | Coat** & feeder | |
| Ca2+, Mg2+ * | Collagenase | Cutting, Scraping, Glass beads | Large | KSR, | MEF | 1,27 |
| Ca2+, Mg2+ * | Free | Cutting | Large | KSR | MEF | 5 |
| Ca2+ *** | Trypsin & Collagenase | Pipetting | Large | KSR | MEF | 2,6,23 |
| Free (EDTA) | Trypsin TrypLE | Pipetting | Single cells, Large | KSR | MEF | 4,5,6,19,28 |
| Free (EDTA) | Free (CDB#) | Pipetting | Single cells | KSR | MEF | 5 |
| Ca2+, Mg2+ * | Dispase, Collagenase TrypLE | Scraping, Pipetting | Large | Free | Mg, Gx, Lm, Vn, Fb, Cg, pVn, pBSP | 8,14,15,20,29 |
| Ca2+, Mg2+ * | Free | Cutting | Large | Free | Vn | 12 |
| Free (EDTA) | Free (CDB#) | Pipetting | Small | Free | Mg, HBP | 8,30 |
| Ca2+ | Free | Pipetting | Large (Small) | Free | Fb | Present study |
*DEME/F12 or ESF solution16 containing Ca2+, Mg2+.
**MEF: mouse embryonic fibroblast feeder cells on gelatin-coated dishes, Mg: Matrigel, Gx: geltrex, Lm: laminin, Vn: vitronectin, Cg: collagen, pVn: vitronectin-derived peptide, pBSP: bone sialoprotein-derived peptide, HBP: heparin-binding peptide.
***Dissociation solution named CTK containing CaCl2, trypsin, collagenase IV, and KSR23. KSR contains many divalent cations.
# CDB: Cell dissociation Buffer (Life Technologies).