Isolation, Cryopreservation, and Characterization of iPSC-Derived Megakaryocytes

Methods Mol Biol. 2021:2180:539-554. doi: 10.1007/978-1-0716-0783-1_27.

Abstract

Current research in the field of transfusion medicine is focused on developing innovative approaches to generate populations of functional megakaryocytes (MKs) ex vivo. This may open perspectives to establish alternative therapies for donor platelet transfusion in the management of thrombocytopenic patients and pave the way for novel regenerative approaches. Efficient cryopreservation techniques can provide the opportunity for long-term storage and accumulation of necessary amounts of MKs in a ready-to-use manner. However, in this case, besides the viability, it is crucial to consider the recovery of functional MK properties after the impact of freezing. In this chapter, the possibility to cryopreserve iPSC-derived MKs is described. In particular, the methods for a comprehensive analysis of phenotypic and functional features of MKs after cryopreservation are proposed. The use of cryopreserved in vitro-produced MKs may benefit to the field of transfusion medicine to overcome the lack of sufficient blood donors.

Keywords: Cryopreservation; Megakaryocytes; Platelets; Polyploidy; Proplatelets.

MeSH terms

  • Animals
  • Blood Platelets / cytology*
  • Blood Platelets / drug effects
  • Cell Culture Techniques / methods*
  • Cell Differentiation
  • Cell Separation / methods*
  • Cryopreservation / methods*
  • Cryoprotective Agents / pharmacology*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Megakaryocytes / cytology*
  • Megakaryocytes / drug effects

Substances

  • Cryoprotective Agents