Cell Physiology Lab, Institute of Biomedical Problems, Russian Academy of Sciences, Khoroshevskoye shosse, 76a, 123007, Moscow, Russia.
Department of Herniology and Plastic Surgery, A. V. Vishnevsky Institute of Surgery, Bolshaya Serpukhovskaya str, 27, 117997, Moscow, Russia.
Cell Tissue Res. 2018 Jun;372(3):523-533. doi: 10.1007/s00441-017-2766-x. Epub 2018 Jan 11.
Umbilical cord blood mononuclear fraction is a valuable source of hematopoietic stem and progenitor cells (CB HSPCs). The rarity of this population is a serious limitation of its application in cell therapy. Ex vivo expansion enables to significantly amplify the number of hematopoietic precursors of different commitment. Here, we expand CB MNCs in co-culture with human adipose tissue-derived stromal cells (ASCs) to enrich HSPCs and describe phenotypic features of newly formed hematopoietic populations. The CD34-HSPCs demonstrated 6-fold enrichment with 9000 CFUs per 50 × 10 HSPCs on average. A part of the floating HSPCs were bearing lineage markers, while others were primitive precursors (CD133/CD34). Among ASC-associated HSPCs, two subsets of cord blood-borne cells were revealed: СD90/СD45 and СD90/СD45. The proportion of CD3/CD8 and NK-T as well as CD25 and HLA-DR Т cells among СD90/СD45 cells was significantly higher compared to MNCs and floating HSPCs. More than 80% of CD45/СD90 HSPCs were identified as late primitive precursors (CD133/CD34). Thus, CB MNC expansion in the presence of ASCs provides the generation of both lineage committed lymphoid progenitors and CD34/CD133 primitive HSPCs. Substantially enriched with primitive precursors, ASC-associated HSPCs could be considered as a perspective tool for a long-term restoration of hematopoiesis in various hematologic disorders.
脐带血单核细胞部分是造血干细胞和祖细胞（CB HSPCs）的有价值来源。该群体的稀有性是其在细胞治疗中应用的严重限制。体外扩增可显著扩增不同定向的造血前体数量。在这里，我们在与人类脂肪组织来源的基质细胞（ASCs）共培养中扩增 CB MNCs，以富集 HSPCs 并描述新形成的造血群体的表型特征。CD34-HSPCs 的平均富集倍数为 6 倍，每 50×10 HSPCs 有 9000 个 CFU。一部分悬浮 HSPCs 携带谱系标记，而另一部分则为原始前体（CD133/CD34）。在 ASC 相关的 HSPCs 中，发现了两种源自脐带血的细胞亚群：CD90/CD45 和 CD90/CD45。与 MNCs 和悬浮 HSPCs 相比，CD3/CD8 和 NK-T 以及 CD25 和 HLA-DR Т 细胞在 CD90/CD45 细胞中的比例明显更高。超过 80%的 CD45/CD90 HSPCs 被鉴定为晚期原始前体（CD133/CD34）。因此，在 ASC 存在的情况下扩增 CB MNC 可产生谱系定向的淋巴祖细胞和 CD34/CD133 原始 HSPCs。原始前体大量富集，ASC 相关的 HSPCs 可被视为各种血液系统疾病中长期恢复造血的有前途的工具。
