Ex Vivo Expansion of Hematopoietic Stem- and Progenitor Cells from Cord Blood in Coculture with Mesenchymal Stroma Cells from Amnion, Chorion, Wharton's Jelly, Amniotic Fluid, Cord Blood, and Bone Marrow
Ex Vivo Expansion of Hematopoietic Stem- and Progenitor Cells from Cord Blood in Coculture with Mesenchymal Stroma Cells from Amnion, Chorion, Wharton's Jelly, Amniotic Fluid, Cord Blood, and Bone Marrow
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DOI：
10.1089/ten.tea.2013.0073
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发表时间：
2013-12-01
影响因子：
4.1
通讯作者：
Weisbach, Volker
中科院分区：
文献类型：
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作者：
Klein, Caroline;Strobel, Julian;Weisbach, Volker
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）的量不足以进行成人同种异体移植。因此，通常需要两个CB单元。与间充质基质细胞（MSC）共培养的CB HSPCs的体外扩增可能是一种替代方法。研究了必须在侵入性程序中获得的骨髓来源的MSC是否可以被来自羊膜、绒毛膜、沃顿氏胶、羊水和CB的MSC替代，所述骨髓来源的MSC引入另外的供体并增加患者的传染性疾病的风险，所述骨髓来源的MSC可以从胎盘组织分离，当CB被采样时，所述胎盘组织是容易获得的。在两步离体共培养中，将来自冷冻保存的CB的单核细胞与不同的MSC饲养层在补充有细胞因子（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）测定以及通过测量CD 34（+）和CD 45（+）细胞来分析扩增率。由于每平方厘米的MSC单层只有5x 10（2）到1x 10（4）个CD 34（+）-细胞，我们观察到了非常高的扩增率，CFU为80到391，000，CD 34（+）-细胞为70到313，000，CD 45（+）-细胞为200到352，000。LTC-IC部分膨胀。与文献相比，我们发现来自所有不同来源的MSC对CD 34（+）-细胞的扩增率更高。这可能是由于我们使用的MSC单层中5x 10（2）至1x 10（+）CD 34-细胞的数量非常少。相比之下，观察到CFU的高扩增率为80至391，000，CD 34（+）-为70至313，000，CD 45（+）-为200至352，000。然而，与来自羊膜、绒毛膜和沃顿氏胶的MSC相比，来自骨髓的MSC对CD 34（+）-细胞的扩增显著更有效。骨髓间充质干细胞与脐血间充质干细胞及羊水间充质干细胞比较无显著性差异。我们得出结论，来自胎盘组织的MSC可能有助于HSPC的扩增，至少如果在扩增培养物中实施低数量的CD 34（+）-细胞/cm（2）MSC-单层和高TPO浓度。
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 5x10(2) to 1x10(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 5x10(2) to 1x10 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.