Dear colleagues!
Does anyone have experience in isolation of non-cultured MSC from bone marrow? Our research group is most interested in patients with primary myelofibrosis. If somebody have such experience, we kindly request to share with us your knowledge.
Please You look at following publication:
Article Gli1 + Mesenchymal Stromal Cells Are a Key Driver of Bone Ma...
https://reader.elsevier.com/reader/sd/pii/S1934590917300796?token=E116F3BF55808522EE5A2845FE98AB2BADDD4B1CC61105A4117C02B5C773B2696BEB1B6E3A72C0B83270D74AB2C3ADEB
Human cell isolation and GANT61 treatmentHuman MSCs (MPN patients or controls from hip replacement surgeries) were isolated by tissue culture plastic adherence after themononuclear cell fraction was separated by Ficoll. Cells were seeded at a density of 5x106cells/flask. MSCs were maintained in me-dium (Mesenpan; PAN Biotech) containing 10% fetal bovine serum (Life Technologies), 1% penicillin-streptomycin-glutamine (LifeTechnologies). Medium was changed every third day and cells were split at near confluence. MSC surface phenotype was assessedat passage 1 using antibodies against the following antigens: CD45 (clone HI30), CD105 (SN6), CD73 (AD2). Antibodies were pur-chased from BD Biosciences (San Jose, USA) or eBiosciences (San Dieg USA). To study the effect of GANT61, cells were treatedfor 24-48h with 10mM GANT61 (Sigma) in DMSO versus vehicle (DMSO).
Can you please provide more detail about your definition of non-cultured and also of MSC?
Why don’t you want to culture? What surface markers are you interested in to define human MSC specifically?
You can extract BM-MSCs from iliac crest of patients, also for characterizing these stem cells first of all you must isolate your target cells by magnetic activated cell sorting (MACS) and using CD markers for example CD90 that specific for BM-MSCs. you can also check our review paper it can help you.
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