Can human hematopoietic stem cells be cultured ex vivo?
Can human hematopoietic stem cells be cultured ex vivo?
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人类造血干细胞可以离体培养吗？
DOI：
10.1002/stem.5530120503
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Verfaillie,CM
中科院分区：
文献类型：
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作者：
Verfaillie,CM
诱导人类造血干细胞增殖的因素尚不明确。需要对此类生长因子进行进一步表征，以开发诱导人类造血干细胞延长增殖和扩增的离体培养系统。能够启动和维持长期培养系统（LTC-IC）的人类或小鼠造血祖细胞代表了非常原始的造血祖细胞群体。我们和其他人已经证明，当与基质层直接接触培养时，可以维持一小部分此类 LTC-IC。此外，补充有二至九种细胞因子的无基质长期培养物可以诱导未成熟的人类造血祖细胞的增殖和分化。然而，在这种培养物中五周后，70-90% 的原始 LTC-IC 就会丢失。我们描述了一种“基质非接触”培养系统，其中祖细胞通过 0.4 μm 微孔膜与基质分开培养，该膜可防止细胞基质接触，但允许扩散因子自由通过。在这种文化中，原始祖细胞不仅可以分化为定型祖细胞，而且比在德克特文化中得到更大程度的维持。我们将讨论 1) 造血祖细胞和骨髓基质之间的直接接触，2) 可溶性基质衍生因子和 3) 先前描述的生长促进和假定的生长抑制细胞因子在 LTC-IC 体外维持和潜在扩展中的相对贡献。
The factors that induce proliferation of the human hematopoietic stem cell are ill defined. Further characterization of such growth factors will be needed to develop ex vivo culture systems that induce prolonged proliferation and expansion of human hematopoietic stem cells. Human or murine hematopoietic progenitors that can initiate and sustain long-term culture systems (LTC-IC) represent a population of very primitive hematopoietic progenitors. When cultured in direct contact with stromal layers, we and others have demonstrated that a fraction of such LTC-IC can be maintained. In addition, stroma-free long-term cultures supplemented with two to nine cytokines can induce proliferation and differentiation of immature human hematopoietic progenitors. However, 70–90% of primitive LTC-IC are lost after five weeks in such cultures. We describe a “stroma-non-contact” culture system, in which progenitors are cultured separated from stroma by a 0.4 μm microporous membrane which prevents cell stroma contact but allows free passage of diffusible factors. Primitive progenitors in such cultures can not only differentiate into committed progenitors but also are maintained to a greater extent than inDextercultures. We will discuss the relative contribution of 1) direct contact between hematopoietic progenitors and bone marrow stroma, 2) soluble stroma-derived factors and 3) previously characterized growth promoting and presumed growth inhibitory cytokines in the in vitro maintenance and potential expansion of LTC-IC.