I have been having some difficulties isolating bone marrow MSC from mouse. Do you generally isolate and culture them under normoxia or hypoxia and how long would this take to reach confluency? there are many contaminating cells too.
Hello
you can culture MSCs by using Alpha MEM+10% ES qualified FBS+2mM Glutamine and Pen/Strep
pleas , see these links , Good Luck
http://www.e-jot.com/article/S2214-031X(14)00085-0/abstract
http://www.nature.com/nprot/journal/v4/n1/full/nprot.2008.221.html
http://www.stemcell.com/~/media/Technical%20Resources/5/28367_mesencult.pdf?la=en
http://www.scielo.br/pdf/aob/v14n1/en_a04v14n1.pdf
http://www.springerprotocols.com/Abstract/doi/10.1007/978-1-62703-128-8_21
http://www.ncbi.nlm.nih.gov/pubmed/19131962
http://www.sciencedirect.com/science/article/pii/S0014482704000084
http://www.ncbi.nlm.nih.gov/pubmed/19089363
it is not a big deal.
sacrifice the mice.
clean the the femur and cut it in two parts.
take 0.5 ml syringe.
flush the bone marrow using MEM medium twice.
collect the cells in 15 ml falcon.
wash twice with mem growth medium.
seed the cells into 90 mm PD and icubate in CO2 incubator for 24-48 hrs.
MSC will settle down and start proliferating and HSC will remain suspended.
Discard the HSC containing medium after 4 days and subculture MSC.
Thank you all for your response regarding my question. Appreciate that.