1 Department of Transfusion Medicine and Hemostaseology, University Hospital of Erlangen-Nuremberg , Erlangen, Germany .
Tissue Eng Part A. 2013 Dec;19(23-24):2577-85. doi: 10.1089/ten.tea.2013.0073.
In most cases, the amount of hematopoietic stem and progenitor cells (HSPCs) in a single cord blood (CB) unit is not sufficient for allogenic transplantation of adults. Therefore, two CB units are usually required. The ex vivo expansion of HSPCs from CB in coculture with mesenchymal stroma cells (MSCs) might be an alternative. It was investigated, whether bone marrow-derived MSCs, which have to be obtained in an invasive procedure, introduce a further donor and increases the risk of transmissible infectious diseases for the patient can be replaced by MSCs from amnion, chorion, Wharton's jelly, amniotic fluid, and CB, which can be isolated from placental tissue which is readily available when CB is sampled. In a two-step ex vivo coculture mononuclear cells from cryopreserved CB were cultured with different MSC-feederlayers in a medium supplemented with cytokines (stem cell factor, thrombopoietin [TPO], and granulocyte colony-stimulating factor). Expansion rates were analyzed as well, by long-term culture-initiating cell (LTC-IC) and colony-forming unit (CFU) assays, as by measuring CD34(+)- and CD45(+)-cells. Due to the comparably low number of 5×10(2) to 1×10(4) CD34(+)-cells per cm(2) MSC-monolayer, we observed comparably high expansion rates from 80 to 391,000 for CFU, 70 to 313,000 for CD34(+)-, and 200 to 352,000 for CD45(+)-cells. Expansion of LTC-IC was partly observed. Compared to the literature, we found a better expansion rate of CD34(+)-cells with MSCs from all different sources. This is probably due to the comparably low number of 5×10(2) to 1×10 CD34(+)-cells per cm(2) MSC-monolayer we used. Comparably, high expansion rates were observed from 80 to 391,000 for CFUs, 70 to 313,000 for CD34(+)-, and 200 to 352,000 for CD45(+)-cells. However, the expansion of CD34(+)-cells was significantly more effective with MSCs from bone marrow compared to MSCs from amnion, chorion, and Wharton's jelly. The comparison of MSCs from bone marrow with MSCs from CB and amniotic fluid showed no significant difference. We conclude that MSCs from placental tissues might be useful in the expansion of HSPCs, at least if low numbers of CD34(+)-cells per cm(2) MSC-monolayer and a high TPO concentration are implemented in the expansion culture.
在大多数情况下，单个脐带血 (CB) 单位中的造血干细胞和祖细胞 (HSPC) 数量不足以进行异基因移植成人。因此，通常需要两个 CB 单位。在与间质基质细胞 (MSCs) 共培养中从 CB 体外扩增 HSPC 可能是一种替代方法。研究了骨髓来源的 MSCs 是否可以替代从羊膜、绒毛膜、沃顿胶、羊水和 CB 中分离的 MSCs，因为骨髓来源的 MSCs需要通过侵入性程序获得，并且可能会增加患者可传播传染病的风险。CB 采样时胎盘组织很容易获得。在两步法体外共培养中，冷冻保存的 CB 单核细胞在添加细胞因子（干细胞因子、血小板生成素[TPO]和粒细胞集落刺激因子）的培养基中与不同的 MSC 饲养层共培养。还通过长期培养起始细胞 (LTC-IC) 和集落形成单位 (CFU) 测定以及测量 CD34(+)和 CD45(+)细胞来分析扩增率。由于 MSC 单层每平方厘米比较低的 5×10(2)到 1×10(4)个 CD34(+)细胞数量，我们观察到 CFU 从 80 到 391,000、CD34(+)从 70 到 313,000 和 CD45(+)从 200 到 352,000 的比较高的扩增率。部分观察到 LTC-IC 的扩增。与文献相比，我们发现来自所有不同来源的 MSCs 的 CD34(+)细胞的扩增率更好。这可能是由于我们使用的 MSC 单层每平方厘米比较低的 5×10(2)到 1×10 CD34(+)细胞数量。类似地，CFU 从 80 到 391,000、CD34(+)从 70 到 313,000 和 CD45(+)从 200 到 352,000 的扩增率也很高。然而，与从羊膜、绒毛膜和沃顿胶中分离的 MSCs 相比，从骨髓中分离的 MSCs 对 CD34(+)细胞的扩增效果更为显著。与从 CB 和羊水分离的 MSCs 相比，从骨髓分离的 MSCs 之间没有显著差异。我们得出结论，胎盘组织来源的 MSCs 可能对 HSPC 的扩增有用，至少在扩增培养中实施每平方厘米 MSC 单层比较低数量的 CD34(+)细胞和高 TPO 浓度时。
