Li Xingfu, Deng Zhenhan, Lu Wei
Department of Orthopedics, Shenzhen Second People's Hospital (The First Affiliated Hospital of Shenzhen University, Health Science Center), Shenzhen, China.
Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Shenzhen University Medical School, Shenzhen, China.
Animal Model Exp Med. 2024 Dec;7(6):793-801. doi: 10.1002/ame2.12515. Epub 2024 Dec 9.
Native cartilage has low capacity for regeneration because it has very few progenitor cells. Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) and human umbilical cord-derived MSCs (hUC-MSCs) have been employed as promising sources of stem cells for cartilage injury repair. Reproduction of hyaline cartilage from MSCs remains a challenging endeavor. The paracrine factors secreted by chondrocytes possess the capability to induce chondrogenesis from MSCs.
The conditioned medium derived from chondrocytes was utilized to induce chondrogenic differentiation of hUCB-MSCs and hUC-MSCs. The expression levels of collagen type I alpha 1 chain (Col1a1), collagen type II alpha 1 chain (Col2a1), and SRY-box transcription factor 9 (SOX9) were assessed through quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB), and immunofluorescence (IF) assays. To elucidate the mechanism of differentiation, the concentration of transforming growth factor-β1 (TGF-β1) in the conditioned medium of chondrocytes was quantified using enzyme-linked immunosorbent assay (ELISA). Meanwhile, the viability of cells was assessed using Cell Counting Kit-8 (CCK-8) assays.
The expression levels of Col2a1 and SOX9 were found to be higher in induced hUC-MSCs compared to those in induced hUCB-MSCs. The conditioned medium of chondrocytes contained TGF-β1. The CCK-8 assays revealed that the proliferation rate of hUC-MSCs was significantly higher compared to that of hUCB-MSCs.
The chondrogenic potential and proliferation capacity of hUC-MSCs surpass those of hUCB-MSCs, thereby establishing hUC-MSCs as a superior source of seed cells for cartilage tissue engineering.
天然软骨的再生能力较低，因为其祖细胞很少。人脐带血间充质干细胞（hUCB-MSCs）和人脐带间充质干细胞（hUC-MSCs）已被用作软骨损伤修复的有前景的干细胞来源。从间充质干细胞再生透明软骨仍然是一项具有挑战性的工作。软骨细胞分泌的旁分泌因子具有诱导间充质干细胞向软骨分化的能力。
利用软骨细胞条件培养基诱导hUCB-MSCs和hUC-MSCs的软骨分化。通过定量实时聚合酶链反应（qRT-PCR）、蛋白质免疫印迹（WB）和免疫荧光（IF）分析评估Ⅰ型胶原α1链（Col1a1）、Ⅱ型胶原α1链（Col2a1）和SRY盒转录因子9（SOX9）的表达水平。为阐明分化机制，采用酶联免疫吸附测定（ELISA）定量软骨细胞条件培养基中转化生长因子-β1（TGF-β1）的浓度。同时，使用细胞计数试剂盒-8（CCK-8）分析评估细胞活力。
发现诱导后的hUC-MSCs中Col2a1和SOX9的表达水平高于诱导后的hUCB-MSCs。软骨细胞条件培养基中含有TGF-β1。CCK-8分析显示，hUC-MSCs的增殖率显著高于hUCB-MSCs。
hUC-MSCs的软骨生成潜力和增殖能力超过hUCB-MSCs，从而确立hUC-MSCs作为软骨组织工程中更优质的种子细胞来源。
