Aleksander-Konert Ewelina, Paduszyński Piotr, Zajdel Alicja, Dzierżewicz Zofia, Wilczok Adam
Department of Biopharmacy, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, ul. Jednosci 8, 41-200 Sosnowiec, Poland.
Department of Health Care, Silesian Medical College, ul. Mickiewicza 29, 40-085 Katowice, Poland.
Cell Mol Biol Lett. 2016 Aug 12;21:11. doi: 10.1186/s11658-016-0016-y. eCollection 2016.
In this study, we evaluated the usefulness of two commercially available hyaluronic acid-based hydrogels, HyStem and HyStem-C, for the cultivation of Wharton's jelly mesenchymal stem cells (WJ-MSCs) and their differentiation towards chondrocytes.
The WJ-MSCs were isolated from umbilical cord Wharton's jelly using the explant method and their immunophenotype was evaluated via flow cytometry analysis. According to the criteria established by the International Society for Cellular Therapy, they were true MSCs. We assessed the ability of the WJ-MSCs and chondrocytes to grow in three-dimensional hydrogels and their metabolic activity. Chondrogenesis of WJ-MSCs in the hydrogels was determined using alcian blue and safranin O staining and real-time PCR evaluation of gene expression in the extracellular matrixes: collagen type I, II, III and aggrecan.
Chondrocytes and WJ-MSCs cultured in the HyStem and HyStem-C hydrogels adopted spherical shapes, which are characteristic for encapsulated cells. The average viability of the WJ-MSCs and chondrocytes in the HyStem hydrogels was approximately 67 % when compared with the viability in 2D culture. Alcian blue and safranin O staining revealed intensive production of proteoglycans by the cells in the HyStem hydrogels. Increased expression of collagen type II and aggrecan in the WJ-MSCs cultured in the HyStem hydrogel in the presence of chondrogenic medium showed that under these conditions, the cells have a high capacity to differentiate towards chondrocytes. The relatively high viability of WJ-MSCs and chondrocytes in both HyStem hydrogels suggests the possibility of their use for chondrogenesis.
The results indicate that WJ-MSCs have some degree of chondrogenic potential in HyStem and HyStem-C hydrogels, showing promise for the engineering of damaged articular cartilage.
在本研究中，我们评估了两种市售的基于透明质酸的水凝胶HyStem和HyStem - C用于培养沃顿胶间充质干细胞（WJ - MSCs）及其向软骨细胞分化的效用。
采用组织块法从脐带沃顿胶中分离WJ - MSCs，并通过流式细胞术分析评估其免疫表型。根据国际细胞治疗协会制定的标准，它们是真正的间充质干细胞。我们评估了WJ - MSCs和软骨细胞在三维水凝胶中的生长能力及其代谢活性。使用阿尔辛蓝和番红O染色以及细胞外基质中基因表达的实时PCR评估来确定水凝胶中WJ - MSCs的软骨形成：I型、II型、III型胶原蛋白和聚集蛋白聚糖。
在HyStem和HyStem - C水凝胶中培养的软骨细胞和WJ - MSCs呈现出球形，这是被包裹细胞的特征。与二维培养中的活力相比，HyStem水凝胶中WJ - MSCs和软骨细胞的平均活力约为67%。阿尔辛蓝和番红O染色显示HyStem水凝胶中的细胞大量产生蛋白聚糖。在软骨形成培养基存在的情况下，在HyStem水凝胶中培养的WJ - MSCs中II型胶原蛋白和聚集蛋白聚糖的表达增加，表明在这些条件下，细胞具有向软骨细胞分化的高能力。两种HyStem水凝胶中WJ - MSCs和软骨细胞相对较高的活力表明它们可用于软骨形成。
结果表明，WJ - MSCs在HyStem和HyStem - C水凝胶中具有一定程度的软骨形成潜力，为受损关节软骨的工程修复展现出前景。
