Augustyniak Ewelina, Suchorska Wiktoria Maria, Trzeciak Tomasz, Richter Magdalena
Radiobiology Laboratory, Greater Poland Cancer Centre, 61‑866 Poznan, Poland.
Department of Orthopedics and Traumatology, Poznan University of Medical Sciences, 61‑545 Poznan, Poland.
Mol Med Rep. 2017 May;15(5):2402-2414. doi: 10.3892/mmr.2017.6335. Epub 2017 Mar 16.
The development of human induced pluripotent stem cells (hiPSCs) is considered a turning point in tissue engineering. However, more data are required to improve understanding of key aspects of the cell differentiation process, including how specific chondrogenic processes affect the gene expression profile of chondrocyte‑like cells and the relative value of cell differentiation markers. The main aims of the present study were as follows: To determine the gene expression profile of chondrogenic‑like cells derived from hiPSCs cultured in mediums conditioned with HC‑402‑05a cells or supplemented with transforming growth factor β3 (TGF‑β3), and to assess the relative utility of the most commonly‑used chondrogenic markers as indicators of cell differentiation. These issues are relevant with regard to the use of human fibroblasts in the reprogramming process to obtain hiPSCs. Human fibroblasts are derived from mesoderm and thus share a wide range of properties with chondrocytes, which originate from the mesenchyme. The hiPSCs were obtained from human primary dermal fibroblasts during a reprogramming process. Two methods, both involving embryoid bodies (EB), were used to obtain chondrocytes from the hiPSCs: EBs formed in the presence of a chondrogenic medium with TGF‑β3 (10 ng/ml) and EBs formed in a medium conditioned with growth factors from HC‑402‑05a cells. Based on reverse transcription-quantitative polymerase chain reaction analysis, the results demonstrated that hiPSCs are capable of effective chondrogenic differentiation, with the cells obtained in the HC‑402‑05a medium presenting with morphological features and markers characteristic of mature human chondrocytes. In contrast, cells differentiated in the presence of TGF‑β3 presented with certain undesirable hypertrophic characteristics. Several genes, most notably runt‑related transcription factor 2, transforming growth factor β2 and transforming growth factor β3, were good markers of advanced and late hiPSC chondrogenic differentiation, whereas transforming growth factor β3I, II, III receptors and bone morphogenetic protein-2, bone morphogenetic protein-4 and growth differentiation factor 5 were less valuable. These findings provide valuable data on the use of stem cells in cartilage tissue regeneration.
人诱导多能干细胞（hiPSCs）的发展被认为是组织工程学中的一个转折点。然而，需要更多数据来增进对细胞分化过程关键方面的理解，包括特定软骨生成过程如何影响类软骨细胞的基因表达谱以及细胞分化标志物的相对价值。本研究的主要目的如下：确定在HC-402-05a细胞条件培养基中培养或添加转化生长因子β3（TGF-β3）的情况下，源自hiPSCs的类软骨细胞的基因表达谱，并评估最常用的软骨生成标志物作为细胞分化指标的相对效用。这些问题与在重编程过程中使用人成纤维细胞来获得hiPSCs有关。人成纤维细胞源自中胚层，因此与源自间充质的软骨细胞具有广泛的共同特性。hiPSCs是在重编程过程中从人原代表皮成纤维细胞获得的。使用两种均涉及胚状体（EB）的方法从hiPSCs中获得软骨细胞：在含有TGF-β3（10 ng/ml）的软骨生成培养基中形成的EB，以及在来自HC-402-05a细胞的生长因子条件培养基中形成的EB。基于逆转录定量聚合酶链反应分析，结果表明hiPSCs能够有效进行软骨生成分化，在HC-402-05a培养基中获得的细胞呈现出成熟人软骨细胞的形态特征和标志物。相比之下，在TGF-β3存在下分化的细胞呈现出某些不良的肥大特征。几个基因，最显著的是 runt相关转录因子2、转化生长因子β2和转化生长因子β3，是hiPSC软骨生成分化晚期的良好标志物，而转化生长因子β3I、II、III受体以及骨形态发生蛋白-2、骨形态发生蛋白-4和生长分化因子5的价值较低。这些发现为干细胞在软骨组织再生中的应用提供了有价值的数据。
