need help for culture protacal of new stem cell - (Jan/29/2006 )

i was familiar with the culture of bone marrow stem cell ,but now my boss ask me prepare the culture of new stem cell ,i need the protacal of new stem cell,are there any body let me share his experience?thank you in advance!

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Hi!

You mean establishment of a new stem cell line ?

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Hi,

Any adult or fetal stem cell (=Mesenchymal stem cell) can be cultured as normal routine culture process.
Use any basal culture medium supllimented with serum and antibiotics. If you can provide the source of stem cell it ma ybe easy to mention particular ingradients. You can proceed the same way you tried with bone marrow stem cells.

Best,
A

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hi... i'm also doing research on stem cells. i think it is da same protocol with other cell culture. but, i'm faced a problem to culture bone marrow. can u tell me how to take bone marrow from rat and how to culture it. thx thx

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I'm not sure from your message whether you are culturing HSCs/MSCs but from what I know of MSC cell culture:

Basically, you can either flush out the rat tibia/femur and collect the cells that way, or you can isolate them using a centrifuge, which appears to be more accurate (and is the way I do it). you cut both ends of the bone off and put in a microfuge tube, supported by inserts (made of cut-up microfuge tubes/pippete tips-whatever you find easiest, to support the bone). you then centrifuge at 2000 rpm for ~5 mins and collect the marrow pellet, before re-suspending in medium and culturing as you would MSCs from human bone.
See link below:

http://www.ncbi.nlm.nih.gov/entrez/query.f...l=pubmed_docsum

However, when culturing rat MSCs as opposed to human, it's best to change the media more frequently, as they grow more rapidly, and note that they also respond better to a lower plating density than human MSCs.

Hope this is helpful.

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Hi Friend,

My boss has asked me to isolate & culture Bone marrow stem cells. I m totally new to this work. can you tell me the protocol please. i m isolating mononuclear cells from Bone marrow by Ficoll density gradient separation and consider that as Stem cell population. Am i right? I dont have magnetic separation or FACS facility. how to be sure that population contain BMSCs?
Pls help me.
Regards
Poonam

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