Could you please share a protocol for culture and expansion of CD34 cells? Also, how to keep them in culture for 10-14 days without differentiation?
Thanks!
I'm going to assume you mean CD34+ hematopoietic stem cells.
Although it is possible to expand hematopoietic progenitors in culture, to do this without any differentiation is not possible. Various hematopoietic growth factors will allow you to support the growth of stem cells into progenitors (KL, FL, TPO, GM-CSF, IL-3, IL-6, G-CSF). Other growth factors can help support various lineages of cells if that is what is desired. Generally, adding more cytokines supports more growth and more differentiation. There are small molecule agents such as the Aryl hydrocarbon receptor antagonist (e.g., StemRegenin 1) that can help minimize differentiation of stem cells. Bone marrow stroma (mesenchymal cells) or endothelial cells have also been shown to help support the maintenance of stem cells. Combinations of all these elements are also possible.
Hello
For more information visit these link
http://www.hindawi.com/journals/bmri/2014/435215/
http://www.bloodjournal.org/content/93/7/2244?sso-checked=true
http://www.stemcell.com/~/media/Technical%20Resources/9/1/1/7/C/29954_TB_StemSpan.pdf
http://www.sciencedirect.com/science/article/pii/S2215017X14000253
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2974206/
Hi
hope this article useful for you
http://www.bloodjournal.org/content/95/1/102.short?sso-checked=true
Best method is to use cytokine combination of FLT3, TPO and SCF and apply co-culture strategy to expand CD34+ cells. Advisable to use cord tissue derived MSCs as a co-culture plastic adherent cell type. You can expect best results with this.
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