Division of Cardiovascular Medicine, Stanford University, 300 Pasteur Drive, Stanford, CA 94305-5406, USA.
Arterioscler Thromb Vasc Biol. 2010 May;30(5):984-91. doi: 10.1161/ATVBAHA.110.202796. Epub 2010 Feb 18.
We examined the effect of delivery modality on the survival, localization, and functional effects of exogenously administered embryonic stem cells (ESCs) or endothelial cells derived from them (ESC-ECs) in the ischemic hindlimb.
Murine ESCs or ESC-ECs were stably transduced with a construct for bioluminescence imaging (BLI) and fluorescent detection. In a syngeneic murine model of limb ischemia, ESCs or ESC-ECs were delivered by intramuscular (IM), intrafemoral artery (IA), or intrafemoral vein injections (n=5 in each group). For 2 weeks, cell survival and localization were tracked by BLI and confirmed by immunohistochemistry, and functional improvement was assessed by laser Doppler perfusion. BLI showed that ESCs localized to the ischemic limb after IM or IA, but not after intrafemoral vein administration. Regardless of the route of administration, ESCs were detected outside the hindlimb circulation in the spleen or lungs. ESCs did not improve limb perfusion and generated teratomas. In contrast, ESC-ECs delivered by all 3 modalities localized to the ischemic limb, as assessed by BLI. Most surprisingly, ESC-EC injected intrafemoral vein eventually localized to the ischemic limb after initially lodging in the pulmonary circulation. Immunohistochemical studies confirmed the engraftment of ESC-ECs into the limb vasculature after 2 weeks. Notably, ESC-ECs were not detected in the spleen or lungs after 2 weeks, regardless of route of administration. Furthermore, ESC-ECs significantly improved limb perfusion and neovascularization compared with the parental ESCs or the vehicle control group.
In contrast to parental ESCs, ESC-ECs preferentially localized in the ischemic hindlimb by IA, IM, and intrafemoral vein delivery. ESC-ECs engrafted into the ischemic microvasculature, enhanced neovascularization, and improved limb perfusion.
我们研究了分娩方式对缺血后肢中外源性给予的胚胎干细胞（ESCs）或其衍生的内皮细胞（ESC-ECs）的存活、定位和功能效果的影响。
将荧光素酶成像（BLI）和荧光检测的构建体稳定转导到小鼠 ESC 或 ESC-EC 中。在同种异体小鼠肢体缺血模型中，通过肌肉内（IM）、股动脉（IA）或股静脉注射（每组 5 只）给予 ESC 或 ESC-EC。在 2 周的时间里，通过 BLI 追踪细胞的存活和定位，并通过免疫组织化学进行确认，通过激光多普勒灌注评估功能改善。BLI 显示 ESC 在 IM 或 IA 后定位于缺血肢体，但在股静脉给药后则不行。无论给药途径如何，ESC 都在脾脏或肺部的下肢循环外被检测到。ESC 并未改善肢体灌注，并且产生了畸胎瘤。相比之下，通过所有 3 种方式给予的 ESC-EC 都能通过 BLI 定位到缺血肢体。最令人惊讶的是，最初在肺循环中定位的 ESC-EC 静脉内注射最终也能定位到缺血肢体。免疫组织化学研究证实了 ESC-EC 在 2 周后定植于肢体血管。值得注意的是，无论给药途径如何，2 周后都未在脾脏或肺部检测到 ESC-EC。此外，与亲代 ESC 或载体对照组相比，ESC-EC 显著改善了肢体灌注和新生血管化。
与亲代 ESC 不同，ESC-EC 优先通过 IA、IM 和股静脉给药定位于缺血后肢。ESC-EC 定植于缺血性微血管，增强了新生血管化并改善了肢体灌注。
