Kabara Maki, Kawabe Jun-ichi, Matsuki Motoki, Hira Yoshiki, Minoshima Akiho, Shimamura Kohei, Yamauchi Atsushi, Aonuma Tatsuya, Nishimura Masato, Saito Yukihiro, Takehara Naofumi, Hasebe Naoyuki
Division of Cardiovascular, Respiratory and Neurology, Department of Medicine, Asahikawa Medical University, Asahikawa, Japan.
Department of Cardiovascular Regeneration and Innovation, Asahikawa Medical University, Asahikawa, Japan.
Lab Invest. 2014 Dec;94(12):1340-54. doi: 10.1038/labinvest.2014.121. Epub 2014 Oct 20.
Adventitial microvessels, vasa vasorum in the vessel walls, have an active role in the vascular remodeling, although its mechanisms are still unclear. It has been reported that microvascular pericytes (PCs) possess mesenchymal plasticity. Therefore, microvessels would serve as a systemic reservoir of stem cells and contribute to the tissues remodeling. However, most aspects of the biology of multipotent PCs (mPCs), in particular of pathological microvessels are still obscure because of the lack of appropriate methods to detect and isolate these cells. In order to examine the characteristics of mPCs, we established immortalized cells residing in adventitial capillary growing at the injured vascular walls. We recently developed in vivo angiogenesis to observe adventitial microvessels using collagen-coated tube (CCT), which also can be used as an adventitial microvessel-rich tissue. By using the CCT, CD146- or NG2-positive cells were isolated from the adventitial microvessels in the injured arteries of mice harboring a temperature-sensitive SV40 T-antigen gene. Several capillary-derived endothelial cells (cECs) and PCs (cPCs) cell lines were established. cECs and cPCs maintain a number of key endothelial and PC features. Co-incubation of cPCs with cECs formed capillary-like structure in Matrigel. Three out of six cPC lines, termed capillary mPCs demonstrated both mesenchymal stem cell- and neuronal stem cell-like phenotypes, differentiating effectively into adipocytes, osteoblasts, as well as schwann cells. mPCs differentiated to ECs and PCs, and formed capillary-like structure on their own. Transplanted DsRed-expressing mPCs were resident in the capillary and muscle fibers and promoted angiogenesis and myogenesis in damaged skeletal muscle. Adventitial mPCs possess transdifferentiation potential with unique phenotypes, including the reconstitution of capillary-like structures. Their phenotype would contribute to the pathological angiogenesis associated with vascular remodeling. These cell lines also provide a reproducible cellular tool for high-throughput studies on angiogenesis, vascular remodeling, and regeneration as well.
血管外膜微血管，即血管壁中的血管滋养管，在血管重塑中发挥着积极作用，尽管其机制尚不清楚。据报道，微血管周细胞（PCs）具有间充质可塑性。因此，微血管将作为干细胞的全身储存库，并有助于组织重塑。然而，由于缺乏检测和分离这些细胞的适当方法，多能PCs（mPCs）生物学的大多数方面，尤其是病理性微血管的方面仍然不清楚。为了研究mPCs的特征，我们建立了驻留在受损血管壁生长的外膜毛细血管中的永生化细胞。我们最近开发了体内血管生成技术，使用胶原包被管（CCT）观察外膜微血管，CCT也可作为富含外膜微血管的组织。通过使用CCT，从携带温度敏感型SV40 T抗原基因的小鼠受伤动脉的外膜微血管中分离出CD146或NG2阳性细胞。建立了几种源自毛细血管的内皮细胞（cECs）和周细胞（cPCs）细胞系。cECs和cPCs保持了许多关键的内皮细胞和PC特征。cPCs与cECs共同孵育在基质胶中形成了毛细血管样结构。六个cPC系中有三个，称为毛细血管mPCs，表现出间充质干细胞和神经干细胞样表型，能有效分化为脂肪细胞、成骨细胞以及雪旺细胞。mPCs分化为内皮细胞和周细胞，并自行形成毛细血管样结构。移植表达DsRed的mPCs驻留在毛细血管和肌纤维中，并促进受损骨骼肌的血管生成和肌生成。外膜mPCs具有独特表型的转分化潜能，包括重建毛细血管样结构。它们的表型将导致与血管重塑相关的病理性血管生成。这些细胞系也为血管生成、血管重塑和再生的高通量研究提供了可重复的细胞工具。
