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•Ex Vivo Expansion Of Hematopoietic Stem Cells Using A Defined Culture Medium

Autologous or allogeneic hematopoietic stem cell (HSC) transplantation is a commonly used therapy for cancer patients following chemotherapy to reconstitute the ablated hematopoietic system. Ex vivo expansion of HSC is frequently exploited to overcome the limited availability of HSCs and progenitors for clinical applications and the understanding of hematopoiesis. The use of conventional basal cell culture medium supplemented with bovine serum often has resulted in unpredictable HSC expansion and presents potential exposure to adventitious viral agents (AVAs). For this reason, we have investigated the development of defined cell culture reagents capable of supporting optimal HSC ex vivo expansion and differentiation. A number of formulations were evaluated for their ability to support survival and expansion of CD34+ HSCs. Granulocyte colony stimulating factor (G-CSF)-mobilized HSCs obtained from healthy individuals (>95% CD34+) were used in this study. All cell culture media were supplemented with recombinant human cytokines Flt-3 ligand, stem cell factor (SCF) and thrombopoietin (TPO) in a 14-day in vitro expansion period. Several developed reagents supported cell expansion 100 - 300 fold after 14 days compared to 150 - 400 fold expansion supported by the currently commercially available media. In analyzing the expanded cultures in the test formulations, it was determined that the majority of cells had undergone differentiation into progenitors of the lineages of myeloid, lymphoid, erythroid and megakaryocyte. Only a small fraction of cells retained the CD34+ phenotype after 14 days. Of interest, one of the developed formulations was capable of expanding and maintaining the CD34+ population to levels comparable or greater than that of commercially available media. In conclusion, preliminary tests of a panel of formulations have confirmed that CD34+ HSCs can be expanded and maintained in a defined medium for up to 14 days, and extensive proliferation and expansion into hematopoietic lineages are achievable with administered cytokine cocktails.

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•Ex Vivo Expansion Of Hematopoietic Stem Cells Using A Defined Culture Medium

SOURCE: SAFC