Kubosch Eva Johanna, Heidt Emanuel, Niemeyer Philipp, Bernstein Anke, Südkamp Norbert P, Schmal Hagen
Department of Orthopaedics and Trauma Surgery, Medical Center, Faculty of Medicine, - Albert-Ludwigs-University of Freiburg, Freiburg, Germany.
Department of Orthopaedics and Traumatology, Odense University Hospital, Odense, Denmark.
Int Orthop. 2017 May;41(5):991-998. doi: 10.1007/s00264-017-3400-y. Epub 2017 Jan 25.
The use of passaged chondrocytes is the current standard for autologous chondrocyte implantation (ACI). De-differentiation due to amplification and donor site morbidity are known drawbacks highlighting the need for alternative cell sources.
Via clinically validated flow cytometry analysis, we compared the expression of human stem cell and cartilage markers (collagen type 2 (Col2), aggrecan (ACAN), CD44) of chondrocytes (CHDR), passaged chondrocytes for ACI (CellGenix™), bone marrow derived mesenchymal stem cells (BMSC), and synovial derived stem cells (SDSC).
Primary, human BMSC and SDSC revealed similar adipogenic, osteogenic, and chondrogenic differentiation potential and stem cell marker expression. However, the expression of the chondrogenic markers Col2 and ACAN was statistically significant higher in SDSC. CHDR and SDSC expressed ACAN and CD44 equally, but Col2 was expressed more strongly on the SDSC surface. The marker expression of SDSC from osteoarthritic joints (Kellgren-Lawrence score ≥3) versus normal knees (Kellgren-Lawrence score ≤2) did not differ. Similarly, there was no difference between temporarily frozen and fresh SDSC. Col2 and ACAN surface expression declined with further passaging, whereas CD44 remained unchanged. We observed the same effect after reducing the serum content. When comparing CHDR for ACI with SDSC of the same passage (P2/3), both Col2 and ACAN, correlating with clinical outcome, were expressed higher in SDSC.
In summary, SDSC demonstrated high differentiation potential and a stable chondrogenic phenotype. They might therefore be better suitable for ACI than BMSC or passaged CHDR.
传代软骨细胞的应用是目前自体软骨细胞植入(ACI)的标准方法。因扩增导致的去分化和供区并发症是已知的缺点,这凸显了对替代细胞来源的需求。
通过临床验证的流式细胞术分析,我们比较了软骨细胞(CHDR)、用于 ACI 的传代软骨细胞(CellGenix™)、骨髓间充质干细胞(BMSC)和滑膜来源干细胞(SDSC)的人类干细胞和软骨标志物(Ⅱ型胶原(Col2)、聚集蛋白聚糖(ACAN)、CD44)的表达。
原代人类 BMSC 和 SDSC 显示出相似的成脂、成骨和软骨分化潜能以及干细胞标志物表达。然而,SDSC 中软骨生成标志物 Col2 和 ACAN 的表达在统计学上显著更高。CHDR 和 SDSC 对 ACAN 和 CD44 的表达相同,但 Col2 在 SDSC 表面表达更强。骨关节炎关节(凯尔格伦 - 劳伦斯评分≥3)与正常膝关节(凯尔格伦 - 劳伦斯评分≤2)的 SDSC 的标志物表达没有差异。同样,临时冷冻和新鲜的 SDSC 之间也没有差异。随着进一步传代,Col2 和 ACAN 的表面表达下降,而 CD44 保持不变。降低血清含量后我们观察到了相同的效果。当将用于 ACI 的 CHDR 与同一代(P2/3)的 SDSC 进行比较时,与临床结果相关的 Col2 和 ACAN 在 SDSC 中的表达更高。
总之,SDSC 表现出高分化潜能和稳定的软骨生成表型。因此,它们可能比 BMSC 或传代 CHDR 更适合用于 ACI。