Rammal H, Beroud J, Gentils M, Labrude P, Menu P, Kerdjoudj H, Velot E
Faculté de Médecine, UMR 7365, CNRS-Université de Lorraine, Biopôle, Vandœuvre-lès-Nancy, France.
Biomed Mater Eng. 2013;23(4):299-309. doi: 10.3233/BME-130754.
Polyelectrolyte multilayer (PEMs) films made of poly(allylamine hydrochloride) (PAH) as polycation and poly(styrene sulfonate) (PSS) as polyanion, with a PAH ending layer, can be used as a coating in order to improve the anti-thrombogenicity and patency of vascular grafts in vascular engineering field. They induce strong adhesion of mature endothelial cells on glass, expanded polytetrafluoroethylene and cryopreserved arteries. Despite their outstanding effect on mature and progenitor endothelial cells, PEMs ending with PAH showed a poor outcome on Wharton's jelly mesenchymal stem cells (WJ-MSCs) culture.
The aim of this work was to examine the influence of the ending charge of PEMs on WJ-MSCs behavior.
WJ-MSCs amplified until the 3rd passage were seeded and cultured on (PAH-PSS)3-PAH and on (PAH-PSS)4 coated glass for 10 days. Stem cell phenotype was checked by flow cytometry and cell morphology was followed by bright field microscopy.
Flow cytometry analysis showed that WJ-MSCs were positive for MSC's markers CD73, CD90 and CD105 and negative for hematopoietic markers CD34 and CD45. Light microscopy showed development of nodule-like structures after 10 days of culture on (PAH-PSS)3-PAH, which resulted in a disturbance of cell monolayer. Whereas WJ-MSCs cultured on (PAH-PSS)4 ending with PSS showed a normal cell growth like on collagen and reached confluence after 10 days.
The culture surface seems to have a determining role in WJ-MSC's "spatial" behavior, which could be considered in the field of tissue engineering.
以聚（烯丙胺盐酸盐）（PAH）作为聚阳离子、聚（苯乙烯磺酸盐）（PSS）作为聚阴离子制成的聚电解质多层（PEMs）薄膜，其末端层为PAH，可作为涂层用于改善血管工程领域中血管移植物的抗血栓形成能力和通畅性。它们能诱导成熟内皮细胞在玻璃、膨体聚四氟乙烯和冷冻保存的动脉上强烈黏附。尽管PEMs对成熟和祖代内皮细胞有显著作用，但以PAH结尾的PEMs在沃顿胶间充质干细胞（WJ-MSCs）培养中效果不佳。
本研究旨在探讨PEMs末端电荷对WJ-MSCs行为的影响。
将扩增至第3代的WJ-MSCs接种于（PAH-PSS）3-PAH和（PAH-PSS）4包被的玻璃上培养10天。通过流式细胞术检测干细胞表型，用明场显微镜观察细胞形态。
流式细胞术分析显示，WJ-MSCs对间充质干细胞标志物CD73、CD90和CD105呈阳性，对造血标志物CD34和CD45呈阴性。光学显微镜显示，在（PAH-PSS）3-PAH上培养10天后出现结节样结构，导致细胞单层紊乱。而在以PSS结尾的（PAH-PSS）4上培养的WJ-MSCs表现出与在胶原蛋白上相似的正常细胞生长，10天后达到汇合。
培养表面似乎在WJ-MSCs的“空间”行为中起决定性作用，这在组织工程领域值得考虑。
