Vitacolonna M, Belharazem D, Hohenberger P, Roessner E D
Division of Surgical Oncology and Thoracic Surgery, Department of Surgery, University Medical Centre Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
Institute of Pathology, University Medical Centre Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
Cell Tissue Bank. 2017 Mar;18(1):27-43. doi: 10.1007/s10561-016-9606-1. Epub 2016 Dec 21.
Transplantation of a cell-seeded graft may improve wound healing after radiotherapy. However, the survival of the seeded cells depends on a rapid vascularization of the graft. Co-culturing of adult stem cells may be a promising strategy to accelerate the vessel formation inside the graft. Thus, we compared the in vivo angiogenic potency of mesenchymal stem cells (MSC) and endothelial progenitor cells (EPC) using dorsal skinfold chambers and intravital microscopy.
Cells were isolated from rat bone marrow and adipose tissue and characterized by immunostaining and flow cytometry. Forty-eight rats received a dorsal skinfold chamber and were divided into 2 main groups, irradiated and non-irradiated. Each of these 2 groups were further subdivided into 4 groups: unseeded matrices, matrices + fibroblasts + pericytes, matrices + fibroblasts + pericytes + MSCs and matrices + fibroblasts + pericytes + EPCs. Vessel densities were quantified semi-automatically using FIJI.
Fibroblasts + pericytes - seeded matrices showed a significantly higher vascular density in all groups with an exception of non-irradiated rats at day 12 compared to unseeded matrices. Co-seeding of MSCs increased vessel densities in both, irradiated and non-irradiated groups. Co-seeding with EPCs did not result in an increase of vascularization in none of the groups.
We demonstrated that the pre-radiation treatment led to a significant decreased vascularization of the implanted grafts. The augmentation of the matrices with fibroblasts and pericytes in co-culture increased the vascularization compared to the non-seeded matrices. A further significant enhancement of vessel ingrowth into the matrices could be achieved by the co-seeding with MSCs in both, irradiated and non-irradiated groups.
接种细胞的移植物移植可能会改善放疗后的伤口愈合。然而，接种细胞的存活取决于移植物的快速血管化。成体干细胞的共培养可能是加速移植物内血管形成的一种有前景的策略。因此，我们使用背皮褶小室和活体显微镜比较了间充质干细胞（MSC）和内皮祖细胞（EPC）的体内血管生成能力。
从大鼠骨髓和脂肪组织中分离细胞，并通过免疫染色和流式细胞术进行表征。48只大鼠接受背皮褶小室，并分为2个主要组，即照射组和未照射组。这2组中的每组又进一步细分为4组：未接种细胞的基质、基质 + 成纤维细胞 + 周细胞、基质 + 成纤维细胞 + 周细胞 + MSC和基质 + 成纤维细胞 + 周细胞 + EPC。使用FIJI半自动定量血管密度。
与未接种细胞的基质相比，接种成纤维细胞 + 周细胞的基质在所有组中均显示出显著更高的血管密度，但在第12天的未照射大鼠组除外。MSC的共接种增加了照射组和未照射组的血管密度。与EPC共接种在任何组中均未导致血管化增加。
我们证明，放疗前治疗导致植入移植物的血管化显著降低。与未接种细胞的基质相比，共培养中用成纤维细胞和周细胞增强基质可增加血管化。在照射组和未照射组中，通过与MSC共接种均可进一步显著增强血管向内生长到基质中。
