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  • 标题:Enrichment of Rabbit Primitive Hematopoietic Cells via MACS Depletion of CD45 + Bone Marrow Cells
  • 本地全文:下载
  • 作者:Jaromír Vašíček ; Andrej Baláži ; Miroslav Bauer
  • 期刊名称:Maine Policy Review
  • 印刷版ISSN:2312-7481
  • 出版年度:2021
  • 卷号:7
  • 期号:1
  • 页码:11
  • DOI:10.3390/magnetochemistry7010011
  • 语种:English
  • 出版社:The University of Maine
  • 摘要:Hematopoietic stem and progenitor cells (HSC/HPCs) of human or few animal species have been studied for over 30 years. However, there is no information about rabbit HSC/HPCs, although they might be a valuable animal model for studying human hematopoietic disorders or could serve as genetic resource for the preservation of animal biodiversity. CD34 marker is commonly used to isolate HSC/HPCs. Due to unavailability of specific anti-rabbit CD34 antibodies, a novel strategy for the isolation and enrichment of rabbit HSC/HPCs was used in this study. Briefly, rabbit bone marrow mononuclear cells (BMMCs) were sorted immunomagnetically in order to remove all mature (CD45<sup>+</sup>) cells. The cells were depleted with overall purity about 60–70% and then cultured in a special medium designed for the expansion of CD34<sup>+</sup> cells. Quantitative Polymerase Chain Reaction (qPCR) analysis confirmed the enrichment of primitive hematopoietic cells, as the expression of CD34 and CD49f increased (<i>p <</i> 0.05) and CD45 decreased (<i>p <</i> 0.001) at the end of culture in comparison to fresh BMMCs. However, cell culture still exhibited the presence of CD45<sup>+</sup> cells, as identified by flow cytometry. After gating on CD45<sup>−</sup> cells the MHCI<sup>+</sup>MHCII<sup>−</sup>CD38<sup>+</sup>CD49f<sup>+</sup>CD90<sup>−</sup>CD117<sup>−</sup> phenotype was observed. In conclusion, rabbit HSC/HPCs might be isolated and enriched by the presented method. However, further optimization is still required.
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