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

  1. Supplement KnockOut™ SR XenoFree Complete Medium with an additional 10% KnockOut™ SR XenoFree (25% final concentration) and 10% dimethyl sulfoxide (DMSO).  
  2. 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).  
  3. Gently resuspend the cell pellet with cryopreservation medium without triturating. 
  4. 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.