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. 2019 Jun 7;14:134–147. doi: 10.1016/j.omtm.2019.05.015

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© 2019 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Media Comparison for CD34⁺ HSPC Culture and Expansion

(A) Experimental scheme. Purified CD34⁺ HSPCs from 2 different HDs were thawed and cultured in X-Vivo 15 (green), SCGM (blue), StemSpan (orange) and HSC Brew (gray) in the presence of SCF (300 ng/mL), Flt3-L (300 ng/mL), and TPO (100 ng/mL). Cultures were analyzed daily by FCM for cell counts and expression of stem cell markers. Cells were additionally subjected to colony-forming unit (CFU) assay on day 3 post-thaw. (B and C) Cell counts assessed with counting beads (B) and percentage of CD34⁺CD90⁺ HSPCs^prim (C), determined by FCM at the indicated time points of culture in the different media. Top: donor A; bottom: donor B. Error bars indicate the mean of 2 independent technical duplicates ± SD; *adjusted p ≤ 0.05, two-way ANOVA with Geisser-Greenhouse correction. (D) Number of BFU-E, CFU-GM, and CFU-GEMM colonies in the CFU assay, determined after 2 weeks of culture in MethoCult. HSC Brew was excluded from CFU assay due to insufficient cell numbers.