Department of Biomaterials, Radboud University Medical Center, Nijmegen, The Netherlands. Department of VIP service, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, People's Republic of China.
Biofabrication. 2014 Mar;6(1):015005. doi: 10.1088/1758-5082/6/1/015005. Epub 2014 Jan 15.
The aim of this study was to comparatively evaluate the angiogenic capacity of cocultures using either human bone marrow- or human adipose tissue-derived mesenchymal stem cells (MSCs) (BM- or AT-MSCs) with human umbilical vein endothelial cells (HUVECs) both in vitro and in vivo at early time points (i.e. days 3 and 7). In vitro, cells were either monocultured (i.e. BM-MSCs, AT-MSCs or HUVECs) or cocultured (i.e. BM-MSCs/HUVECs and AT-MSCs/HUVECs) on Thermanox® (2-dimensional, 2D) or in collagen gels (3-dimensional, 3D). For the in vivo experiment, cells (cocultures) were embedded in collagen gels and implanted subcutaneously in nude mice. For both in vitro and in vivo experiments, samples were collected on days 3 and 7 and histologically processed for hematoxylin-eosin and platelet endothelial cell adhesion molecule (PECAM-1; CD31) staining. For in vivo samples, quantitative parameters for evaluating angiogenesis included CD31-positive staining percentage, total vessel-like structure (VLS) area percentage, VLS density, and average VLS area (i.e. the size of per VLS). In vitro results showed the formation of VLS in both cocultures, while none of the monocultures showed VLS formation, irrespective of 2D or 3D culture condition. Although VLS formation occurred after in vivo implantation, no significant difference in angiogenic capacity was observed between the two cocultures, either on day 3 or on day 7. Further, VLS density decreased and anastomosis of the new human vessels with the murine host vasculature occurred over time. In conclusion, this study demonstrated that AT-MSCs/HUVECs and BM-MSCs/HUVECs have equal angiogenic capacity both in vitro and in vivo, and that vessels from donor origin can anastomose with the host vasculature within seven days of implantation.
本研究旨在比较评估人骨髓间充质干细胞（BM-MSCs）或人脂肪组织来源的间充质干细胞（AT-MSCs）与脐静脉内皮细胞（HUVECs）的共培养物在体外和早期时间点（即第 3 天和第 7 天）的血管生成能力。在体外，细胞分别进行单培养（即 BM-MSCs、AT-MSCs 或 HUVECs）或共培养（即 BM-MSCs/HUVECs 和 AT-MSCs/HUVECs），分别在 Thermanox®（二维，2D）或胶原凝胶（三维，3D）上进行。对于体内实验，将细胞（共培养物）嵌入胶原凝胶中，并植入裸鼠皮下。对于体外和体内实验，在第 3 天和第 7 天收集样本，并进行苏木精-伊红和血小板内皮细胞黏附分子（PECAM-1；CD31）染色的组织学处理。对于体内样本，评估血管生成的定量参数包括 CD31 阳性染色百分比、总血管样结构（VLS）面积百分比、VLS 密度和平均 VLS 面积（即每个 VLS 的大小）。体外结果显示两种共培养物中均形成了 VLS，而无论 2D 或 3D 培养条件如何，单培养物均未形成 VLS。尽管在体内植入后发生了 VLS 形成，但在第 3 天或第 7 天，两种共培养物的血管生成能力均无显著差异。此外，VLS 密度降低，新的人血管与宿主血管吻合。总之，本研究表明，AT-MSCs/HUVECs 和 BM-MSCs/HUVECs 在体外和体内均具有同等的血管生成能力，并且供体来源的血管可以在植入后 7 天内与宿主血管吻合。
