Suchorska Wiktoria Maria, Augustyniak Ewelina, Richter Magdalena, Łukjanow Magdalena, Filas Violetta, Kaczmarczyk Jacek, Trzeciak Tomasz
Radiobiology Lab, Greater Poland Cancer Centre, Poznan, Poland.
Department of Orthopaedics and Traumatology, Poznan University of Medical Sciences, Poland.
Postepy Hig Med Dosw (Online). 2017 Jun 19;71(0):500-509. doi: 10.5604/01.3001.0010.3831.
Human articular cartilage has a poor regenerative capacity. This often results in the serious joint disease- osteoarthritis (OA) that is characterized by cartilage degradation. An inability to self-repair provided extensive studies on AC regeneration. The cell-based cartilage tissue engineering is a promising approach for cartilage regeneration. So far, numerous cell types have been reported to show chondrogenic potential, among others human embryonic stem cells (hESCs).
However, the currently used methods for directed differentiation of human ESCs into chondrocyte-like cells via embryoid body (EB) formation, micromass culture (MC) and pellet culture (PC) are not highly efficient and require further improvement. In the present study, these three methods for hESCs differentiation into chondrocyte-like cells in the presence of chondrogenic medium supplemented with diverse combination of growth factors (GFs) were evaluated and modified.
The protocols established here allow highly efficient, simple and inexpensive production of a large number of chondrocyte-like cells suitable for transplantation into the sites of cartilage injury. The most crucial issue is the selection of appropriate GFs in defined concentration. The obtained stem-derived cells reveal the presence of chondrogenic markers such as type II collagen, Sox6 and Sox9 as well as the lack or significantly lower level of pluripotency markers including Nanog and Oct3/4.
The most efficient method is the differentiation throughout embryoid bodies. In turn, chondrogenic differentiation via pellet culture is the most promising method for implementation on clinical scale. The most useful GFs are TGF-β1, -3 and BMP-2 that possess the most chondrogenic potential. These methods can also be used to obtain chondrocyte-like cells from differentiating induced pluripotent stem cells (iPSCs).
人类关节软骨的再生能力较差。这常常导致以软骨退化为特征的严重关节疾病——骨关节炎（OA）。软骨无法自我修复引发了对软骨再生的广泛研究。基于细胞的软骨组织工程是一种很有前景的软骨再生方法。到目前为止，已报道许多细胞类型具有软骨形成潜力，其中包括人类胚胎干细胞（hESCs）。
然而，目前通过胚胎体（EB）形成、微团培养（MC）和微珠培养（PC）将人类胚胎干细胞定向分化为软骨样细胞的方法效率不高，需要进一步改进。在本研究中，对这三种在添加不同生长因子（GFs）组合的软骨形成培养基存在下将人类胚胎干细胞分化为软骨样细胞的方法进行了评估和改进。
这里建立的方案能够高效、简单且低成本地大量生产适合移植到软骨损伤部位的软骨样细胞。最关键的问题是选择确定浓度的合适生长因子。所获得的干细胞来源细胞显示出软骨形成标志物如II型胶原蛋白、Sox6和Sox9的存在，以及包括Nanog和Oct3/4在内的多能性标志物的缺失或显著较低水平。
最有效的方法是通过胚胎体进行分化。反过来，通过微珠培养进行软骨形成分化是在临床规模上实施最有前景的方法。最有用的生长因子是具有最大软骨形成潜力的TGF-β1、-3和BMP-2。这些方法也可用于从诱导多能干细胞（iPSCs）分化中获得软骨样细胞。
