Xenobiotic kidney organogenesis from human mesenchymal stem cells using a growing rodent embryo
Xenobiotic kidney organogenesis from human mesenchymal stem cells using a growing rodent embryo
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DOI：
10.1681/asn.2005101043
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发表时间：
2006-04-01
影响因子：
13.6
通讯作者：
Kobayashi, Eiji
中科院分区：
文献类型：
--
作者：
Yokoo, Takashi;Fukui, Akira;Kobayashi, Eiji
鉴于同种异体器官移植的局限性，最终的治疗方案是从自体干细胞中建立自体器官，并将其作为同系移植物移植回供体患者体内。据报道，在生长的啮齿动物胚胎中培养的人间充质干细胞（hMSC）可以在空间和时间适当的发育环境中分化，促进肾发生的第一步。作为走向临床应用的另一步，该系统被修改为进展到完整的功能器官发生。从子宫分离大鼠胚胎（E11.5），并将骨髓来源的hMSC（用胶质细胞系衍生的神经营养因子腺病毒转染和用LacZ逆转录病毒转染）植入肾发生部位。48小时后，输尿管芽伸长，完成了最初的分支。切取后肾，体外器官培养24 h，移植于单侧肾切除大鼠网膜，生长2周。它们增大并表现出正常的肾脏结构和超微结构。在整个再生肾中鉴定出hMSC衍生的LacZ阳性细胞，并且在形态上与驻留肾细胞相同。将发育中的后肾移植到LacZ转基因大鼠中显示，新肾血管系统起源于宿主循环。最后，从扩张的输尿管收集液体，并测量尿素氮和肌酐。与移植大鼠血清相比，这些液体中的水平高得多（分别为840.3 +/- 184.6 vs 30.4 +/- 10.8和10.1 +/- 3.1 vs 0.3 +/- 0.2 mg），表明新肾可能产生尿液。总之，这些发现表明hMSC可以分化成成熟的肾脏结构，具有替代丧失的肾功能的潜力。
Given the limits of allogenic organ transplantation, an ultimate therapeutic solution is to establish a self-organ from autologous stem cells and transplant them as syngrafts back into donor patients. It was reported previously that human mesenchymal stem cells (hMSC) that are cultivated in growing rodent embryos can differentiate within a spatially and temporally appropriate developmental milieu, facilitating the first step of nephrogenesis. As another step toward clinical application, the system was modified for progression to complete functional organogenesis. Rat embryos (E11.5) were isolated from uteri, and bone marrow-derived hMSC, which were transfected adenovirally with glial cell line-derived neurotrophic factor and retrovirally with LacZ, were implanted into the nephrogenic site. Forty-eight hours later, ureteric buds were elongated and initial branching was completed. The metanephroi were dissected out, developed further using in vitro organ culture for 24 h, transplanted into the omentum of a uninephrectomized rat, and grown for 2 wk. They enlarged and exhibited normal kidney structure and ultrastructure. hMSC-derived LacZ-positive cells were identified throughout the regenerated kidney and were morphologically identical to resident renal cells. Transplantation of developing metanephroi into the LacZ transgenic rat revealed that neo-kidney vasculature originated from the host circulation. Finally, fluid was collected from expanded ureters, and urea nitrogen and creatinine were measured. Levels were much higher in these fluids compared with transplanted rat sera (840.3 +/- 184.6 versus 30.4 +/- 10.8 and 10.1 +/- 3.1 versus 0.3 +/- 0.2 mg, respectively), suggesting that the neo-kidney may produce urine. Taken together, these findings suggest that hMSC can differentiate into a mature renal structure with the potential to replace lost kidney function.