Expandable Megakaryocyte Cell Lines Enable Clinically Applicable Generation of Platelets from Human Induced Pluripotent Stem Cells
Expandable Megakaryocyte Cell Lines Enable Clinically Applicable Generation of Platelets from Human Induced Pluripotent Stem Cells
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DOI：
10.1016/j.stem.2014.01.011
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发表时间：
2014-04-03
期刊：
影响因子：
23.9
通讯作者：
Eto, Koji
中科院分区：
文献类型：
--
作者：
Nakamura, Sou;Takayama, Naoya;Eto, Koji
依赖于捐献者的血小板供应常常不足以满足输血需求。为了解决这个问题，我们开发了一种临床上适用的策略，用于从人多能干细胞（PSC）中衍生功能性血小板。该方法涉及通过BMI 1和BCL-XL的过表达以分别抑制衰老和凋亡以及c-MYC的受限过表达以促进增殖，从PSC衍生的造血祖细胞建立稳定的永生化巨核细胞祖细胞系（imMKCL）。即使在冷冻保存后，所得的imMKCL也可以在培养物中延长时间（4-5个月）进行扩增。在生长的imMKCL中停止c-MYC、BMI 1和BCL-XL的过表达导致产生CD 42 b（+）血小板，其功能性基于一系列体外和体内测定与天然血小板相当。稳健的扩增能力和有效的血小板产生的组合意味着适当选择的imMKCL克隆代表了用于临床应用的hPSC衍生的血小板的潜在取之不尽的来源。
The donor-dependent supply of platelets is frequently insufficient to meet transfusion needs. To address this issue, we developed a clinically applicable strategy for the derivation of functional platelets from human pluripotent stem cells (PSCs). This approach involves the establishment of stable immortalized megakaryocyte progenitor cell lines (imMKCLs) from PSC-derived hematopoietic progenitors through the overexpression of BMI1 and BCL-XL to respectively suppress senescence and apoptosis and the constrained overexpression of c-MYC to promote proliferation. The resulting imMKCLs can be expanded in culture over extended periods (4-5 months), even after cryopreservation. Halting the overexpression of c-MYC, BMI1, and BCL-XL in growing imMKCLs led to the production of CD42b(+) platelets with functionality comparable to that of native platelets on the basis of a range of assays in vitro and in vivo. The combination of robust expansion capacity and efficient platelet production means that appropriately selected imMKCL clones represent a potentially inexhaustible source of hPSC-derived platelets for clinical application.