Chondrogenic commitment of human umbilical cord blood and umbilical cord-derived mesenchymal stem cells induced by the supernatant of chondrocytes: A comparison study

Q1 Health Professions

Animal models and experimental medicine Pub Date : 2024-12-09 DOI:10.1002/ame2.12515

Xingfu Li, Zhenhan Deng, Wei Lu

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Abstract

Background

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.

Methods

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.

Results

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.

Conclusions

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.

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软骨细胞上清诱导人脐带血和脐带间充质干细胞的成软骨承诺：一项比较研究。

背景：原生软骨的再生能力较低，因为它的祖细胞很少。人脐带血来源间充质干细胞（hub -MSCs）和人脐带来源间充质干细胞（hUC-MSCs）已被用作软骨损伤修复的有希望的干细胞来源。从间充质干细胞中复制透明软骨仍然是一项具有挑战性的努力。软骨细胞分泌的旁分泌因子具有诱导间充质干细胞成软骨的能力。方法：利用软骨细胞衍生的条件培养基诱导hub - mscs和hUC-MSCs成软骨分化。通过定量实时聚合酶链式反应（qRT-PCR）、免疫印迹（WB）和免疫荧光（IF）检测I型胶原α 1链（Col1a1）、II型胶原α 1链（Col2a1）和SRY-box转录因子9 （SOX9）的表达水平。为阐明分化机制，采用酶联免疫吸附法（ELISA）定量测定软骨细胞条件培养基中转化生长因子-β1 （TGF-β1）的浓度。同时，采用细胞计数试剂盒-8 （CCK-8）检测细胞活力。结果：Col2a1和SOX9在诱导的hUC-MSCs中的表达水平高于诱导的hub - mscs。软骨细胞条件培养基中含有TGF-β1。CCK-8检测结果显示，hUC-MSCs的增殖率明显高于hub - mscs。结论：hUC-MSCs的成软骨潜能和增殖能力超过hub - mscs，从而使hUC-MSCs成为软骨组织工程种子细胞的优越来源。

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