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从腘窝囊肿中分离并鉴定人滑液来源的间充质基质细胞

Isolation and Characterization of Human Synovial Fluid-Derived Mesenchymal Stromal Cells from Popliteal Cyst.

作者信息

Li Fang, Chen Jianglin, Gong Mengjia, Bi Yang, Hu Chengchen, Zhang Yuanyuan, Li Ming

机构信息

Department of Orthopedics, Ministry of Education Key Laboratory of Child Development and Disorders, National Clinical Research Center for Child Health and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children's Hospital of Chongqing Medical University, Chongqing, China.

Department of Pediatric Research Institute, Chongqing Key Laboratory of Pediatrics, Children's Hospital of Chongqing Medical University, Chongqing, China.

出版信息

Stem Cells Int. 2020 Sep 18;2020:7416493. doi: 10.1155/2020/7416493. eCollection 2020.

Abstract

Mesenchymal stem cells (MSCs) are multipotent progenitor cells in adult tissues. The aim of this study is to isolate and identify synovial fluid-derived mesenchymal stromal cells (SF-MSCs) from the popliteal cyst fluid of pediatric patients. SF-MSCs were collected from the popliteal cyst fluid of pediatric patients during cystectomy surgery. After cyst fluid extraction and adherent culturing, in vitro morphology, growth curve, and cell cycle were observed. The expression of stem cell surface markers was analyzed by flow cytometry, and expression of cell marker protein was detected by immunofluorescence. SF-MSCs were cultured in osteogenic, adipogenic, and chondrogenic differentiation medium. The differentiation potential of SF-MSCs was analyzed by alkaline phosphatase (Alizarin Red), Oil Red O, and Alcian blue. Antibody detection of human angiogenesis-related proteins was performed compared with bone marrow mesenchymal stem cells (BM-MSCs). The results show that SF-MSCs from the popliteal cyst fluid of pediatric patients showed a shuttle appearance and logarithmic growth. Flow cytometry analysis revealed that SF-MSCs were negative for hematopoietic lineage markers (CD34, CD45) and positive for MSC markers (CD44, CD73, CD90, and CD105). Interstitial cell marker (vimentin) and myofibroblast-like cell marker alpha-smooth muscle actin (-SMA) were positive. These cells could differentiate into osteogenic, adipogenic, and chondrogenic lineages, respectively. Several types of human angiogenesis-related proteins were detected in the cell secretory fluid. These results show that we successfully obtained SF-MSCs from the popliteal cyst fluid of pediatric patients, which have the potential to be a valuable source of MSCs.

摘要

间充质干细胞(MSCs)是成体组织中的多能祖细胞。本研究的目的是从儿科患者的腘窝囊肿液中分离并鉴定滑膜液源性间充质基质细胞(SF-MSCs)。在囊肿切除手术期间从儿科患者的腘窝囊肿液中收集SF-MSCs。在提取囊肿液并进行贴壁培养后,观察其体外形态、生长曲线和细胞周期。通过流式细胞术分析干细胞表面标志物的表达,并通过免疫荧光检测细胞标志物蛋白的表达。将SF-MSCs培养在成骨、成脂和软骨分化培养基中。通过碱性磷酸酶(茜素红)、油红O和阿尔辛蓝分析SF-MSCs的分化潜能。与骨髓间充质干细胞(BM-MSCs)相比,进行了人类血管生成相关蛋白的抗体检测。结果显示,来自儿科患者腘窝囊肿液的SF-MSCs呈梭形外观并呈对数生长。流式细胞术分析显示,SF-MSCs对造血谱系标志物(CD34、CD45)呈阴性,对MSC标志物(CD44、CD73、CD90和CD105)呈阳性。间质细胞标志物(波形蛋白)和成肌纤维细胞样细胞标志物α-平滑肌肌动蛋白(α-SMA)呈阳性。这些细胞可分别分化为成骨、成脂和软骨谱系。在细胞分泌液中检测到几种类型的人类血管生成相关蛋白。这些结果表明,我们成功地从儿科患者的腘窝囊肿液中获得了SF-MSCs,其有可能成为MSCs的宝贵来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6011/7519976/084a8d149dd0/SCI2020-7416493.001.jpg

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