Isolation and enrichment of rat mesenchymal stem cells (MSCS) and separation of single-colony derived MSCs

Linxia Zhang, Christina Chan

Research output: Contribution to journalArticle

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Abstract

MSCs are a population of adult stem cells that is a promising source for therapeutic applications. These cells can be isolated from the bone marrow and can be easily separated from the hematopoietic stem cells (HSCs) due to their plastic adherence. This protocol describes how to isolate MSCs from rat femurs and tibias. The isolated cells were further enriched against two MSCs surface markers CD54 and CD90 by magnetic cell sorting. Expression of surface markers CD54 and CD90 were then confirmed by flow cytometry analysis. HSC marker CD45 was also included to check if the sorted MSCs were depleted of HSCs. MSCs are naturally quite heterogeneous. There are subpopulations of cells that have different shapes, proliferation and differentiation abilities. These subpopulations all express the known MSCs markers and no unique marker has yet been identified for the different subpopulations. Therefore, an alternative approach to separate out the different subpopulations is using cloning cylinders to separate out single-colony derived cells. The cells derived from the single-colonies can then be cultured and evaluated separately.

LanguageEnglish (US)
Article numbere1852
JournalJournal of Visualized Experiments
Issue number37
DOIs
StatePublished - Mar 2010

Profile

Cell Separation
Stem cells
Mesenchymal Stromal Cells
Rats
Hematopoietic Stem Cells
Cells
Flow cytometry
Cloning
Sorting
Bone
Adult Stem Cells
Plastics
Tibia
Femur
Organism Cloning
Flow Cytometry
Bone Marrow
Population

Keywords

  • Cellular biology
  • Cloning cylinder
  • Flow cytometry
  • Issue 37
  • Magnetic cell sorting
  • Mesenchymal stem cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

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