Koch Jill M, D'Souza Saritha S, Schwahn Denise J, Dixon Ian, Hacker Timothy A
Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA.
Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, USA.
Cytotherapy. 2016 Feb;18(2):219-28. doi: 10.1016/j.jcyt.2015.10.013. Epub 2015 Dec 28.
Existing treatments have limited success in modifying the course of peripheral artery disease, which may eventually lead to limb-threatening ulcers and amputation. Cellular therapies have the potential to provide a new treatment option for this condition, but isolation of cells by conventional means has limitations with respect to reproducibility and scalability.
Induced pluripotent stem cells (iPSCs) were differentiated into precursor cells known as mesenchymoangioblasts (MCAs) and subsequently into mesenchymal stromal cells (MSCs). Hindlimb ischemia in mice was created by ligating both the iliac and femoral arteries of one hindlimb. Immediately after surgery, each animal received intramuscular injections of 5 × 10(6) cells or media in the ischemic limb. Toe necrosis was assessed visually, and hindlimb blood flow was measured by laser Doppler using a set region of interest (ROI) and by tracing the entire foot. Myofiber heterogeneity, nuclear centralization, fatty degeneration, fibrosis and capillary angiogenesis in the gastrocnemius muscle were assessed histologically.
Blood flow in the MCA-derived MSC-treated animals was higher at each day (P <0.006), and these mice recovered faster than control animals (3.6 vs. 2.5 for set ROI; 7.5 vs. 4.1 foot tracing; slope; P <0.001). There was significantly less myofiber heterogeneity, nuclear centralization, fatty degeneration and fibrosis in MCA-derived MSC-treated animals, indicating less tissue damage.
MCA-derived MSCs improved limb blood flow, reduced necrosis and maintained muscle mass and gross muscle appearance. We conclude that MCA-derived MSCs have a significant and protective effect against ischemic insults.
现有治疗方法在改变外周动脉疾病病程方面成效有限，外周动脉疾病最终可能导致肢体溃疡和截肢。细胞疗法有可能为这种疾病提供新的治疗选择，但通过传统方法分离细胞在可重复性和可扩展性方面存在局限性。
将诱导多能干细胞（iPSC）分化为称为间充质血管母细胞（MCA）的前体细胞，随后再分化为间充质基质细胞（MSC）。通过结扎一侧后肢的髂动脉和股动脉来制造小鼠后肢缺血模型。手术后立即给每只动物在缺血肢体进行肌肉注射5×10⁶个细胞或培养基。通过肉眼评估趾坏死情况，并使用设定的感兴趣区域（ROI）通过激光多普勒测量后肢血流量，以及通过描绘整个足部来测量。对腓肠肌中的肌纤维异质性、核中心化、脂肪变性、纤维化和毛细血管生成进行组织学评估。
接受MCA来源的MSC治疗的动物在每一天的血流量都更高（P<0.006），并且这些小鼠比对照动物恢复得更快（设定ROI时分别为3.6对2.5；足部描绘时分别为7.5对4.1；斜率；P<0.001）。接受MCA来源的MSC治疗的动物中肌纤维异质性、核中心化、脂肪变性和纤维化明显更少，表明组织损伤更小。
MCA来源的MSC改善了肢体血流量，减少了坏死，并维持了肌肉质量和肌肉大体外观。我们得出结论，MCA来源的MSC对缺血性损伤具有显著的保护作用。
