Evaluation of human platelet lysate and dimethyl sulfoxide as cryoprotectants for the cryopreservation of human adipose-derived stem cells

Biochem Biophys Res Commun. 2017 Sep 9;491(1):198-203. doi: 10.1016/j.bbrc.2017.07.076. Epub 2017 Jul 13.

Abstract

Cryopreservation provides an effective technique to maintain the functional properties of human adipose-derived stem cells (ASCs). Dimethylsulfoxide (DMSO) and fetal bovine serum (FBS) are frequently used as cryoprotectants for this purpose. However, the use of DMSO can result in adverse effects and toxic reactions and FBS can introduce risks of viral, prion, zoonose contaminations and evoke immune responses after injection. It is therefore crucial to reduce DMSO concentrations and use serum-free solution in the cryopreservation process. Human platelet lysate (PL) is a promising candidate for use as an alternative to DMSO and FBS. Therefore, in this study, with an aim to identify a cryoprotective agent for ASC cryopreservation, we determined the viability, proliferation potential, phenotype, and differentiation potential of fresh ASCs and ASCs cryopreserved using different combinations of three cryoprotective agents: fetal bovine serum (FBS), dimethylsulfoxide (DMSO), and human platelet lysate (PL). The viability of the ASCs cryopreserved with 90% FBS and 10% DMSO, 95% FBS and 5% DMSO, and 97% PL and 3% DMSO was >80%, and the proliferation potentials, cell phenotypes, and differentiation potentials of these groups were similar to those of fresh ASCs. Together, our findings suggest that a combination of 97% PL and 3% DMSO is an ideal cryoprotective agent for the efficient cryopreservation of human ASCs.

Keywords: Adipose-derived stem cells; Cryopreservation; Cryoprotective agent; Platelet lysate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / drug effects
  • Blood Platelets / chemistry*
  • Cell Differentiation / drug effects
  • Cell Extracts / pharmacology
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Cryopreservation / methods*
  • Cryoprotective Agents / pharmacology
  • Dimethyl Sulfoxide / pharmacology*
  • Humans
  • Stem Cells / chemistry
  • Stem Cells / cytology*
  • Stem Cells / drug effects*

Substances

  • Cell Extracts
  • Cryoprotective Agents
  • Dimethyl Sulfoxide