Cell Systems Tips and Tricks
Tip #1: To facilitate cell binding with CELLstart™ substrate—dilute in DPBS that contains calcium and magnesium (Invitrogen part# 14040). CELLstart™ can be used with:
- Human embryonic stem cells (hESCs)
- Mesenchymal stem cells
- Neural stem cells
- Human induced pluripotent stem cells (iPSCs)
- Human feeder cells (e.g., foreskin fibroblasts)
Tip #2: Geltrex Basement Membrane Matrix for cell invasion assays—apply a thick layer (50–200 µL/cm2) onto the growth surface. Incubate for 30 min and you’re good to go!
Tip #3: Cryopreserve hESCs with KnockOut™ SR XenoFree
- Supplement KnockOut™ SR XenoFree Complete Medium with an additional 10% KnockOut™ SR XenoFree (25% final concentration) and 10% dimethyl sulfoxide (DMSO).
- Dilute hESCs at a higher concentration than would normally be passaged (if 1:5 is a routine passage, use 1:3 or 1:4 for freezing).
- Gently resuspend the cell pellet with cryopreservation medium without triturating.
- After dispensing, gently invert to mix the cells.
For best results, hESC vials should be cryopreserved using a controlled-rate freezer (e.g., Thermo Scientific CryoMed Freezer or Nalgene 5200 Cryo 1°C “Mr. Frosty” Freezing Container). Expect some cell death upon recovery.
Tip #4: Use StemPro® MSC SFM XenoFree for serum-free and xeno-free growth of human mesenchymal stem cells (MSCs) and adipose-derived stem cells (ADSCs)—StemPro® MSC SFM XenoFree has also been shown to support the growth of cord blood–derived MSCs, umbilical cord matrix (Wharton’s jelly) MSCs, and human fibroblasts.
Tip #5: Increase AlgiMatrix® 3D Culture System firmness with AlgiMatrix® Firming Buffer—AlgiMatrix® Firming Bufffer is an isotonic, neutral-pH buffered solution designed to increase the firmness of AlgiMatrix® 3D bioscaffolds. Firmer sponges are less susceptible to pore expansion and contraction by cellular and environmental forces, and can also be easily picked up and manipulated with sterile forceps. For a firmer sponge, add AlgiMatrix® Firming Buffer to cells and culture medium prior to adding the cells to the sponge.