Harmouch C, El-Omar R, Labrude P, Decot V, Menu P, Kerdjoudj H
UMR 7365 CNRS-Université de Lorraine, MoPA, Vandæuvre-les-Nancy, France Reviva Regenerative Medicine Center, Middle East Institute of Health, Bsalim, Lebanon.
Biomed Mater Eng. 2013;23(4):273-80. doi: 10.3233/BME-130751.
Mesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several lineages with valuable applications in regenerative medicine. MSCs differentiation is highly dependent on physicochemical properties of the culture substrate, cell density and on culture medium composition.
In this study, we assessed the influence of fetal bovine serum (FBS) level on Wharton's jelly (WJ)-MSCs behavior seeded on polyelectrolyte multilayer films (PEMF) made of four bilayers of poly-allylamine hydrochloride (PAH) as polycation and poly-styrene sulfonate (PSS) as polyanion.
MSCs isolated from WJ by explants method were amplified until the third passage. Their phenotypic characterization was performed by flow cytometry analyses. MSCs were seeded on PEMF, in Endothelial growth medium-2 (EGM-2) supplemented by either 5% or 2% FBS. Cell's behavior was monitored for 20 days by optical microscopy and immunofluorescence.
Until 2 weeks on glass slides, no difference was observed whatever the FBS percentage. Then with 5% FBS, MSCs formed three-dimensional spheroids on PSS/PAH after 20 days of culture with a nuclear aggregate. Whereas, with 2% FBS, these spheroids did not appear and cells grown in 2D conserved the fibroblast-like morphology.
The decrease of FBS percentage from 5% to 2% avoids 3D cell spheroids formation on PAH/PSS. Such results could guide bioengineering towards building 2D structures like cell layers or 3D structures by increasing the osteogenic or chondrogenic differentiation potential of MSCs.
间充质干细胞（MSCs）是多能细胞，能够分化为多个谱系，在再生医学中具有重要应用价值。MSCs的分化高度依赖于培养底物的物理化学性质、细胞密度以及培养基组成。
在本研究中，我们评估了胎牛血清（FBS）水平对接种于由四层聚阳离子聚烯丙胺盐酸盐（PAH）和聚阴离子聚苯乙烯磺酸盐（PSS）制成的聚电解质多层膜（PEMF）上的沃顿胶（WJ）-MSCs行为的影响。
采用组织块法从WJ中分离出MSCs，并扩增至第三代。通过流式细胞术分析对其进行表型鉴定。将MSCs接种于PEMF上，置于添加5%或2% FBS的内皮细胞生长培养基-2（EGM-2）中。通过光学显微镜和免疫荧光监测细胞行为20天。
在载玻片上培养2周内，无论FBS百分比如何，均未观察到差异。然后，在5% FBS条件下，培养20天后，MSCs在PSS/PAH上形成三维球体，伴有核聚集。而在2% FBS条件下，未出现这些球体，细胞在二维平面生长并保持成纤维细胞样形态。
FBS百分比从5%降至2%可避免在PAH/PSS上形成三维细胞球体。这些结果可为生物工程提供指导，通过提高MSCs的成骨或成软骨分化潜能来构建细胞层等二维结构或三维结构。
