Isolation and Culture of Mouse Mesenchymal Stem Cells (MSCs)

 

1. Sacrifice 6-10 weeks old mice by CO2.

2. Isolate the bones and flush out bone marrow with a 5 ml syringe through a 25 gauge needle

    with antibiotics enriched medium

3. transfer the marrow cell suspension to six well plates at a concentration of 1.5x106 nucleated cells per cm2

4. Culture the cell in low glucose DMEM with 10% FBS.

5. After 72 hours, remove the nonadherent cells and renew half of the medium.

6. Trypsinize, harvest, and passage the cells, once the adherent cells reach 70-80% confluence.

7. Passage 3-5 homogenous cells can be used for other experiment.

 

Protocols

 

 
 
 
 
 
 
 

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Ch 1.General Lab Techniques

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Ch 3.DNA and RNA

Ch 4.Genetics

Ch 5.PCR Serials

Ch 6.Protein

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Ch 8.Immunohistoch / immunology

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Ch 10.GC/MS, NMR and Proteomics

Ch 11.Animal Experiments

Ch 12.Worm: C. Elegans

Ch 13.HPLC and TLC

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