Andreeva Elena R, Andrianova Irina V, Sotnezova Elena V, Buravkov Sergey V, Bobyleva Polina I, Romanov Yury A, Buravkova Ludmila B
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia.
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia; Institute of Experimental Cardiology, Cardiology Research Center, Moscow, Russia.
PLoS One. 2015 Apr 28;10(4):e0124939. doi: 10.1371/journal.pone.0124939. eCollection 2014.
The optimisation of haematopoietic stem and progenitor cell expansion is on demand in modern cell therapy. In this work, haematopoietic stem/progenitor cells (HSPCs) have been selected from unmanipulated cord blood mononuclear cells (cbMNCs) due to adhesion to human adipose-tissue derived stromal cells (ASCs) under standard (20%) and tissue-related (5%) oxygen. ASCs efficiently maintained viability and supported further HSPC expansion at 20% and 5% O2. During co-culture with ASCs, a new floating population of differently committed HSPCs (HSPCs-1) grew. This suspension was enriched with СD34+ cells up to 6 (20% O2) and 8 (5% O2) times. Functional analysis of HSPCs-1 revealed cobble-stone area forming cells (CAFCs) and lineage-restricted colony-forming cells (CFCs). The number of CFCs was 1.6 times higher at tissue-related O2, than in standard cultivation (20% O2). This increase was related to a rise in the number of multipotent precursors - BFU-E, CFU-GEMM and CFU-GM. These changes were at least partly ensured by the increased concentration of MCP-1 and IL-8 at 5% O2. In summary, our data demonstrated that human ASCs enables the selection of functionally active HSPCs from unfractionated cbMNCs, the further expansion of which without exogenous cytokines provides enrichment with CD34+ cells. ASCs efficiently support the viability and proliferation of cord blood haematopoietic progenitors of different commitment at standard and tissue-related O2 levels at the expense of direct and paracrine cell-to-cell interactions.
现代细胞治疗对造血干细胞和祖细胞扩增的优化有需求。在这项工作中，造血干细胞/祖细胞（HSPCs）是从未经处理的脐带血单个核细胞（cbMNCs）中筛选出来的，因为它们在标准（20%）和组织相关（5%）氧气条件下能黏附于人脂肪组织来源的基质细胞（ASCs）。ASCs在20%和5%氧气条件下能有效维持细胞活力并支持HSPCs的进一步扩增。在与ASCs共培养期间，出现了一群新的具有不同分化方向的悬浮HSPCs（HSPCs-1）。该悬浮液中CD34+细胞富集了6倍（20%氧气）和8倍（5%氧气）。对HSPCs-1的功能分析显示有鹅卵石区域形成细胞（CAFCs）和谱系限制集落形成细胞（CFCs）。在组织相关氧气条件下CFCs的数量比标准培养（20%氧气）时高1.6倍。这种增加与多能前体细胞——爆式红系集落形成单位（BFU-E）、粒-红-巨噬-巨核集落形成单位（CFU-GEMM）和粒-巨噬集落形成单位（CFU-GM）数量的增加有关。这些变化至少部分是由5%氧气条件下MCP-1和IL-8浓度的增加所保证的。总之，我们的数据表明人ASCs能够从未经分离的cbMNCs中筛选出功能活跃的HSPCs，在不添加外源性细胞因子的情况下对其进一步扩增可实现CD34+细胞的富集。ASCs通过直接和旁分泌的细胞间相互作用，在标准和组织相关氧气水平下有效支持不同分化方向的脐带血造血祖细胞的活力和增殖。
