Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells as an Individual-Specific and Renewable Source of Adult Stem Cells

Methods Mol Biol. 2017:1553:183-190. doi: 10.1007/978-1-4939-6756-8_14.

Abstract

This chapter deals with the employment of human-induced pluripotent stem cells (hiPSCs) as a candidate to differentiate into mesenchymal stem cells (MSCs). This would enable to help establish a regular source of human MSCs with the aim of avoiding the problems associated with procuring the MSCs either from different healthy individuals or patients, limited extraction potentials, batch-to-batch variations or from diverse sources such as bone marrow or adipose tissue. The procedures described herein allow for a guided and ensured approach for the regular maintenance of hiPSCs and their subsequent differentiation into MSCs using the prescribed medium. Subsequently, an easy protocol for the successive isolation and purification of the hiPSC-differentiated MSCs is outlined, which is carried out through passaging and can be further sorted through flow cytometry. Further, the maintenance and expansion of the resultant hiPSC-differentiated MSCs using appropriate characterization techniques, i.e., Reverse-transcription PCR and immunostaining is also elaborated. The course of action has been deliberated keeping in mind the awareness and the requisites available to even beginner researchers who mostly have access to regular consumables and medium components found in the general laboratory.

Keywords: Differentiation; Induced pluripotent stem cells; Mesenchymal stem cells; Non-variable MSC source; Regenerative medicine.

MeSH terms

  • Biomarkers
  • Cell Culture Techniques
  • Cell Differentiation*
  • Cell Proliferation
  • Cells, Cultured
  • Cellular Reprogramming
  • Humans
  • Immunophenotyping
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism*
  • Mesenchymal Stem Cells / cytology*
  • Microscopy, Fluorescence
  • Phenotype
  • Regenerative Medicine

Substances

  • Biomarkers