I have been trying to keep CD34+ HSC undifferentiated in suspension culture, and compared some culture conditions with different cytokine combinations, etc. I also tried the co-culture on primary hMSC but due to donor to donor difference of MSC, I decided to try a mesenchymal cell line, SCP-1. Since SCP-1 does not have contact inhibition, I did mitotic inactivation by both irradiation and mitomycin-C, yet in both cases the feeder layer did not support cobblestone formation in LTC-IC assay (cultured in myelocult media with hydrocortisone). I wonder if using some cytokines or a different medium would help at all. Has anyone have experience with this?
Have you checked out the following reference:? Boiron JM, Dazey B, Cailliot C, Launay B, Attal M, Mazurier F, McNiece IK, Ivanovic Z, Caraux J, Marit G, et al. Large-scale expansion and transplantation of CD34(+) hematopoietic cells: in vitro and in vivo confirmation of neutropenia abrogation related to the expansion process without impairment of the long-term engraftment capacity. Transfusion. 2006;46:1934–1942.
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