Influence of Factors of Cryopreservation and Hypothermic Storage on Survival and Functional Parameters of Multipotent Stromal Cells of Placental Origin

PLoS One. 2015 Oct 2;10(10):e0139834. doi: 10.1371/journal.pone.0139834. eCollection 2015.

Abstract

Human placenta is a highly perspective source of multipotent stromal cells (MSCs) both for the purposes of patient specific auto-banking and allogeneic application in regenerative medicine. Implementation of new GMP standards into clinical practice enforces the search for relevant methods of cryopreservation and short-term hypothermic storage of placental MSCs. In this paper we analyze the effect of different temperature regimes and individual components of cryoprotective media on viability, metabolic and culture properties of placental MSCs. We demonstrate (I) the possibility of short-term hypothermic storage of these cells; (II) determine DMSO and propanediol as the most appropriate cryoprotective agents; (III) show the possibility of application of volume expanders (plasma substituting solutions based on dextran or polyvinylpyrrolidone); (IV) reveal the priority of ionic composition over the serum content in cryopreservation media; (V) determine a cooling rate of 1°C/min down to -40°C followed by immersion into liquid nitrogen as the optimal cryopreservation regime for this type of cells. This study demonstrates perspectives for creation of new defined cryopreservation methods towards GMP standards.

MeSH terms

  • Cell Survival / drug effects*
  • Cells, Cultured
  • Cryopreservation / methods
  • Cryoprotective Agents / pharmacology*
  • Dimethyl Sulfoxide / pharmacology
  • Female
  • Humans
  • Multipotent Stem Cells / drug effects*
  • Placenta / drug effects*
  • Pregnancy

Substances

  • Cryoprotective Agents
  • Dimethyl Sulfoxide

Grants and funding

Funding institution: Bundesministerium für Bildung und Forschung (Federal Ministry of Education and Research, Germany). Funding Program: Richtlinien zur Förderung der Wissenschaftlich-Technischen Zusammenarbeit (WTZ) mit der Ukraine. Funding Program URL: http://www.bmbf.de/foerderungen/20159.php. Project funding reference number: 01DK13013. Project title: Genetische Modifikation und Langzeitlagerung von plazentalen mesenchymalen Stammzellen für klinische Anwendungen (Genetic Modification and long term Storage of Placental derived Mesenchymal Stem cells for clinical applications). Funded authors: DP VP O. Pogozhykh TM O. Prokopyuk. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.