Cryopreserved CD90+ cells obtained from mobilized peripheral blood in sheep: a new source of mesenchymal stem cells for preclinical applications

Cell Tissue Bank. 2016 Mar;17(1):137-45. doi: 10.1007/s10561-015-9526-5. Epub 2015 Jul 29.

Abstract

Mobilized peripheral blood (MPB) bone marrow cells possess the potential to differentiate into a variety of mesenchymal tissue types and offer a source of easy access for obtaining stem cells for the development of experimental models with applications in tissue engineering. In the present work, we aimed to isolate by magnetic activated cell sorting CD90+ cells from MPB by means of the administration of Granulocyte-Colony Stimulating Factor and to evaluate cell proliferation capacity, after thawing of the in vitro culture of this population of mesenchymal stem cells (MSCs) in sheep. We obtained a median of 8.2 ± 0.6 million of CD90+ cells from the 20-mL MPB sample. After thawing, at day 15 under in vitro culture, the mean CD90+ cells determined by flow cytometry was 92.92 ± 1.29 % and cell duplication time determined by crystal violet staining was 47.59 h. This study describes for the first time the isolation, characterization, and post-in vitro culture thawing of CD90+ MSCs from mobilized peripheral blood in sheep. This population can be considered as a source of MSCs for experimental models in tissue engineering research.

Keywords: Cryopreserved CD90+ cells; Mesenchymal stem cells; Mobilized peripheral blood.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Proliferation
  • Cell Separation
  • Cell Shape
  • Cryopreservation / methods*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Immunophenotyping
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Peripheral Blood Stem Cells / cytology*
  • Sheep
  • Thy-1 Antigens / metabolism*

Substances

  • Thy-1 Antigens