Isolation and cultivation of bone marrow-derived mesenchymal stromal cells

Commonly used acronym: BM-MSC

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
bone marrow-derived mesenchymal stromal cells


Method keywords
  • Stem cells
  • stem cell culture
  • bone marrow
  • stem cell isolation
  • mesenchymal stromal cells
Scientific area keywords
  • mesenchymal stromal cells
  • stem cell culture
  • stem cell isolation
Method description

Mononuclear cells (MNC) are isolated from bone marrow aspirates by density gradient centrifugation and washed in Hank's buffered salt solution. MNC are seeded at a cell density of 2 × 10E4 cells/cm2 in low glucose DMEM supplemented with 15% (v/v) heat-inactivated FBS, 2 mM L-glutamine and 0.5% (v/v) antibiotic/antimycotic solution. Cells are incubated at 37°C in a 5% (v/v) CO2-enriched humidified atmosphere, cultured up to 90% confluency, trypsinized, centrifuged, and subcultured at a lower density (5 × 10E3 cells/cm2) for all subsequent passages for 2 weeks.

Lab equipment
  • Biosafety cabinet level 2;
  • Cell incubator;
  • Centrifuge.
Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential


Robust protocol for isolation of bone marrow-derived mesenchymal stromal cells.

References, associated documents and other information


De Kock J, Najar M, Bolleyn J, Al Battah F, Rodrigues RM, Buyl K, Raicevic G, Govaere O, Branson S, Meganathan K, Gaspar JA, Roskams T, Sachinidis A, Lagneaux L, Vanhaecke T, Rogiers V. (2012) Mesoderm-derived stem cells: the link between the transcriptome and their differentiation potential. Stem Cells Dev. 21(18):3309-23

Contact person

Joery De Kock


Vrije Universiteit Brussel (VUB)
Pharmaceutical and Pharmacological Sciences
In Vitro Toxicology and Dermato-Cosmetology
Brussels Region