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利用低氧和碱性成纤维细胞生长因子-2增强人骨髓基质细胞的增殖并优化培养条件

Enhancing proliferation and optimizing the culture condition for human bone marrow stromal cells using hypoxia and fibroblast growth factor-2.

作者信息

Lee Jung-Seok, Kim Seul Ki, Jung Byung-Joo, Choi Seong-Bok, Choi Eun-Young, Kim Chang-Sung

机构信息

Department of Periodontology, Research Institute for Periodontal Regeneration, College of Dentistry, Yonsei University, Seoul, Republic of Korea.

Department of Neurosurgery, Naeun Hospital, Seoul, Republic of Korea.

出版信息

Stem Cell Res. 2018 Apr;28:87-95. doi: 10.1016/j.scr.2018.01.010. Epub 2018 Jan 9.

Abstract

This study aimed to determine the cellular characteristics and behaviors of human bone marrow stromal cells (hBMSCs) expanded in media in a hypoxic or normoxic condition and with or without fibroblast growth factor-2 (FGF-2) treatment. hBMSCs isolated from the vertebral body and expanded in these four groups were evaluated for cellular proliferation/migration, colony-forming units, cell-surface characterization, in vitro differentiation, in vivo transplantation, and gene expression. Culturing hBMSCs using a particular environmental factor (hypoxia) and with the addition of FGF-2 increased the cellular proliferation rate while enhancing the regenerative potential, modulated the multipotency-related processes (enhanced chondrogenesis-related processes/osteogenesis, but reduced adipogenesis), and increased cellular migration and collagen formation. The gene expression levels in the experimental samples showed activation of the hypoxia-inducible factor-1 pathway and glycolysis in the hypoxic condition, with this not being affected by the addition of FGF-2. The concurrent application of hypoxia and FGF-2 could provide a favorable condition for culturing hBMSCs to be used in clinical applications associated with bone tissue engineering, due to the enhancement of cellular proliferation and regenerative potential.

摘要

本研究旨在确定在低氧或常氧条件下、添加或不添加成纤维细胞生长因子-2(FGF-2)的培养基中扩增的人骨髓间充质干细胞(hBMSC)的细胞特性和行为。对从椎体分离并在这四组条件下扩增的hBMSC进行细胞增殖/迁移、集落形成单位、细胞表面特征、体外分化、体内移植及基因表达的评估。利用特定环境因素(低氧)并添加FGF-2培养hBMSC,可提高细胞增殖率,增强再生潜能,调节多能性相关过程(增强软骨生成相关过程/成骨作用,但减少脂肪生成),并增加细胞迁移和胶原蛋白形成。实验样本中的基因表达水平显示,在低氧条件下缺氧诱导因子-1途径和糖酵解被激活,添加FGF-2对此无影响。由于细胞增殖和再生潜能增强,低氧和FGF-2的同时应用可为培养用于骨组织工程相关临床应用的hBMSC提供有利条件。

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