Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, Center of Regenerative Therapies, Hohe Straße 6, Dresden, Saxony, 01069, Germany.
Trinity Centre for Bioengineering, Trinity Biomedical Science Institute, Trinity College Dublin 2, Dublin, Ireland.
Sci Rep. 2017 Jul 25;7(1):6377. doi: 10.1038/s41598-017-06898-2.
Limbal stromal cells (LSCs) from the human ocular surface display mesenchymal stromal cell characteristics in vitro. In this study, we isolated cells from the porcine limbal stroma (pLSCs), characterised them, and evaluated their ability to support angiogenesis and the culture of porcine limbal epithelial stem cells (pLESCs). The isolated cells adhered to plastic and grew in monolayers in vitro using serum-supplemented or serum-free medium. The pLSCs demonstrated expression of CD29, and cross-reactivity with anti-human CD45, CD90, CD105, CD146, and HLA-ABC. However, expression of CD105, CD146 and HLA-ABC reduced when cultured in serum-free medium. PLSCs did not undergo adipogenic or osteogenic differentiation, but differentiated towards the chondrogenic lineage. Isolated cells were also co-cultured with human umbilical vein endothelial cells (HUVECs) in star-shaped Poly(ethylene glycol) (starPEG)-heparin hydrogels to assess their pericyte capacity which supported angiogenesis networks of HUVECs. PLSCs supported the three dimensional HUVEC network for 7 days. The isolated cells were further growth-arrested and evaluated as feeder cells for pLESC expansion on silk fibroin membranes, as a potential carrier material for transplantation. PLSCs supported the growth of pLESCs comparably to murine 3T3 cells. In conclusion, although pLSCs were not completely comparable to their human counterpart, they display several mesenchymal-like characteristics in vitro.
人眼表面的角膜缘基质细胞（LSCs）在体外具有间充质基质细胞的特征。在本研究中，我们从猪角膜缘基质（pLSCs）中分离细胞，对其进行鉴定，并评估其支持血管生成和培养猪角膜缘上皮干细胞（pLESCs）的能力。分离的细胞在体外附着于塑料并在含有血清或无血清培养基的单层中生长。pLSCs 表达 CD29，并与抗人 CD45、CD90、CD105、CD146 和 HLA-ABC 发生交叉反应。然而，在无血清培养基中培养时，CD105、CD146 和 HLA-ABC 的表达减少。PLSCs 不会发生成脂或成骨分化，但向软骨谱系分化。分离的细胞还与人脐静脉内皮细胞（HUVECs）在星形聚乙二醇（starPEG）-肝素水凝胶中共培养，以评估其周细胞能力，从而支持 HUVECs 的血管生成网络。PLSCs 支持 HUVEC 网络三维生长 7 天。进一步对分离的细胞进行生长抑制，并在丝素蛋白膜上作为 pLESC 扩增的饲养细胞进行评估，作为移植的潜在载体材料。PLSCs 支持 pLESCs 的生长与鼠 3T3 细胞相当。总之，尽管 pLSCs 与人类细胞不完全可比，但它们在体外显示出几种间充质样特征。
