Isolation and cultivation of bone marrow-derived mesenchymal stromal cells
Commonly used acronym: BM-MSC
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
Description
- Stem cells
- stem cell culture
- bone marrow
- stem cell isolation
- mesenchymal stromal cells
- mesenchymal stromal cells
- stem cell culture
- stem cell isolation
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.
- Biosafety cabinet level 2;
- Cell incubator;
- Centrifuge.
- 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 KockOrganisations
Vrije Universiteit Brussel (VUB)Pharmaceutical and Pharmacological Sciences
In Vitro Toxicology and Dermato-Cosmetology
Belgium
Brussels Region