By Michal Amit PhD, Joseph Itskovitz-Eldor MD DSc (auth.), Michal Amit, Joseph Itskovitz-Eldor (eds.)
Human pluripotent stem cells, together with human embryonic stem cells and prompted pluripotent stem cells, are a key concentration of present biomedical study. The emergence of state-of-the-art culturing innovations is selling the conclusion of the total power of pluripotent stem cells in uncomplicated and translational learn and in cell-based remedies. This entire and authoritative atlas summarizes greater than a decade of expertise amassed by means of a number one study crew during this box. Hands-on step by step advice for the derivation and culturing of human pluripotent stem cells in outlined stipulations (animal product-free, serum-free, feeder-free) and in non-adhesion suspension tradition are supplied, in addition to tools for analyzing pluripotency (embryoid physique and teratoma formation) and karyotype balance.
The Atlas of Human Pluripotent Stem Cells - Derivation and Culturing will function a reference and consultant to validated researchers and people wishing to go into the promising box of pluripotent stem cells.
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Additional info for Atlas of Human Pluripotent Stem Cells: Derivation and Culturing
Centrifuge for 3 min at 80 × g at a recommended temperature of 4°C. 7. Remove the supernatant and re-suspend the cells gently in 2 ml medium. 8. Place the cell suspension in one well of a 6-well plate, or of a 4-well plate, covered with feeder cells (see Sect. 5). If the thawing procedure succeeds, small colonies should appear 1 day postthawing (Fig. 12). With very good recovery, as demonstrated in Fig. 13, colonies will continue to grow, and will not differ from post-splitting cells. However, in some cases, when either freezing or thawing fails, the cells might not survive, as can be seen in the examples illustrated in Fig.
B) A colony of hESCs CL1 cultured with mTeSR™ containing undifferentiated-like cells with spaces between them. The cell size is larger than usual, and the colony is not round. It seems that there are still some feeder cells at the culture (an example is marked with an arrow). 2 cultured with NutriStem™ with elongated shape. (d) A large colony of hESCs CL1 mainly containing undifferentiated cells. At the colony center, there is an area (marked with a circle) where the cell morphology is unclear.
Bar 100 mM Fig. 17 Colonies from hESC H9 5 days post-thawing. (a) Cells are already differentiated (marked with arrow), differentiation probably occurred upon passage to freezing. (b, c) Undifferentiated colonies with good recovery and size. 3 Methods 33 Fig. 2 hESCs, with disorganized morphology and some apoptotic cells. (a, b, d) Undifferentiated colonies with apoptotic cells (marked by arrow) detaching from the colony. In most cases, apoptotic cells will disappear after a few days, and the surviving cells will remain unchanged.