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体外软骨细胞分化的评估:沙特阿拉伯王国阿卜杜勒阿齐兹国王大学的一项干细胞研究计划。

Evaluation of in vitro chondrocytic differentiation: A stem cell research initiative at the King Abdulaziz University, Kingdom of Saudi Arabia.

作者信息

Al-Yamani Aisha, Kalamegam Gauthaman, Ahmed Farid, Abbas Mohammed, Sait Khalid Hussein Wali, Anfinan Nisreen, Al-Wasiyah Mohammad Khalid, Huwait Etimad A, Gari Mamdouh, Al-Qahtani Mohammed

机构信息

Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia.

Stem Cell Unit, Centre of Excellence in Genomic Medicine Research, King Abdulaziz University, Jeddah, Saudi Arabia.

出版信息

Bioinformation. 2018 Feb 28;14(2):53-59. doi: 10.6026/97320630014053. eCollection 2018.

Abstract

Mesenchymal stem cells (MSCs) from various sources have been used in cartilage differentiation with variable success. Therefore, it is of interest to evaluate the in vitro differentiation potential of the hWJSCs derived from the human umbilical cords into chondrocytes at the stem cell research facility at the King Abdulaziz University. hWJSCs are an attractive choice for tissue engineering and regenerative medical applications including cartilage regeneration. We evaluated the hWJSCs using classical histological and cartilage related gene expression studies. Some of the known parameters were re-examined for consistency at the current laboratory conditions. Early passages (P1-P4) showed short fibroblastic morphology and high expression of MSC related surface markers namely CD29 (99.9%), CD44 (97.8%), CD73 (99.6%), CD90 (95.1%) and CD105 (98.9%). MTT assay showed time dependent increase in hWJSCs proliferation by 61.06% and 206.31% at 48h and 72h respectively. Toluidine blue histology showed that hWJSCs were successfully differentiated into chondrocytes in chondrocytic differentiation medium for 21 days. Differentiated hWJSCs also showed significantly increased expression of collagen type II, aggrecan and SOX9 compared to the undifferentiated control. It should be noted that the determination of the average cell yield, the population doubling time and histological staining wtih alcian blue and/or safronin O is required in future studies for improved evaluation of differentiation. Painless derivation, abundance of stem cells that are hypo-immunogenic and safety issues makes this method advantages to MSCs derived from other sources.

摘要

来自不同来源的间充质干细胞(MSCs)已被用于软骨分化,但成功率各不相同。因此,在阿卜杜勒阿齐兹国王大学的干细胞研究设施中,评估源自人脐带的人脐带华通氏胶间充质干细胞(hWJSCs)在体外向软骨细胞分化的潜力具有重要意义。hWJSCs是组织工程和再生医学应用(包括软骨再生)的一个有吸引力的选择。我们使用经典的组织学和软骨相关基因表达研究来评估hWJSCs。在当前实验室条件下,重新检查了一些已知参数的一致性。早期传代(P1-P4)显示出短纤维细胞形态,并且与MSC相关的表面标志物即CD29(99.9%)、CD44(97.8%)、CD73(99.6%)、CD90(95.1%)和CD105(98.9%)的表达较高。MTT分析显示,hWJSCs在48小时和72小时时的增殖分别随时间增加了61.06%和206.31%。甲苯胺蓝组织学显示,hWJSCs在软骨细胞分化培养基中成功分化为软骨细胞达21天。与未分化对照相比,分化后的hWJSCs还显示出II型胶原蛋白、聚集蛋白聚糖和SOX9的表达显著增加。需要注意的是,在未来的研究中,为了更好地评估分化,需要测定平均细胞产量、群体倍增时间以及用阿尔新蓝和/或番红O进行组织学染色。无痛取材、丰富的低免疫原性干细胞以及安全性问题使得这种方法比源自其他来源的MSCs更具优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/502f/5879944/8b164613b09d/97320630014053F1.jpg

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