Human Umbilical Cord Mesenchymal Stromal Cell Isolation, Expansion, Cryopreservation, and Characterization

Curr Protoc Stem Cell Biol. 2017 May 16:41:1F.18.1-1F.18.23. doi: 10.1002/cpsc.24.

Abstract

Revised methods to derive, expand, and characterize mesenchymal stromal cells (MSCs) from the umbilical cord are provided. Several considerations are taken for GMP compliance including using a closed system isolation method and eliminating several xenogenic components. With this method cells are isolated using mechanical and enzymatic digestion and then expanded with high viabilities that retain >90% viability after cryopreservation. Lastly, characterization methods have been optimized to identify these cells as MSCs according to the ISCT minimal criteria. This method standardizes the process for isolating, expanding, cryopreserving, and characterizing MSCs from the umbilical cord. © 2017 by John Wiley & Sons, Inc.

Keywords: characterization; cryopreservation; culture; isolation; mesenchymal stromal cells (MSCs); platelet lysate; stem cells; umbilical cord (UC).

MeSH terms

  • Adipogenesis
  • Cell Count
  • Cell Proliferation
  • Cell Separation / methods*
  • Cells, Cultured
  • Chondrogenesis
  • Colony-Forming Units Assay
  • Cryopreservation / methods*
  • Flow Cytometry
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Osteogenesis
  • Staining and Labeling
  • Umbilical Cord / cytology*