Multipotential Differentiation of Human Anulus Fibrosus Cells An in Vitro Study
Multipotential Differentiation of Human Anulus Fibrosus Cells An in Vitro Study
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DOI：
10.2106/jbjs.h.01672
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发表时间：
2010-03-01
影响因子：
5.3
通讯作者：
Li, Xudong
中科院分区：
文献类型：
--
作者：
Feng, Gang;Yang, Xinlin;Li, Xudong
背景资料：在椎间盘退变过程中纤维软骨、骨样组织、神经和血管在纤维环中的存在已被充分证明。已经假设分化细胞从椎间盘外迁移是这些组织形成的可能机制。我们假设，正常的纤维环组织中含有多能祖细胞，这是能够分化成软骨和/或纤维软骨细胞，成骨细胞，神经元，血管cells.Methods：我们分离纤维环细胞从非退行性椎间盘的青少年（13至16岁）特发性脊柱侧凸患者和培养细胞在体外诱导培养基中含有不同的刺激。通过流式细胞术进行细胞表面标志物的免疫表型分析。使用免疫染色、细胞组织学染色和逆转录-聚合酶链反应测定脂肪形成、骨形成、软骨形成、神经形成和分化为内皮谱系的标志物的表达。纤维环细胞表达几种有时与间充质干细胞相关的细胞表面抗原，包括CD 29、CD 49 e、CD 51、CD 73、CD 90、CD 105、CD 166、CD 184和Stro-1，以及两种神经干细胞标记物巢蛋白和神经元特异性烯醇化酶。此外，不同的刺激物加入到诱导培养基确定是否纤维环细胞分化成脂肪细胞，成骨细胞，软骨细胞，神经元，或endothelial cells.Conclusions：纤维环细胞分离从非退行性椎间盘可以分化成脂肪细胞，成骨细胞，软骨细胞，神经元，和内皮细胞在体外。
Background: The existence of fibrocartilage, bone-like tissues, nerves, and blood vessels in the anulus fibrosus during intervertebral disc degeneration has been well documented. Migration of differentiated cells from outside the intervertebral disc has been hypothesized as a possible mechanism for the formation of these tissues. We hypothesized that the normal anulus fibrosus tissue contains multipotent progenitor cells, which are able to differentiate into cartilage and/or fibrocartilage cells, osteoblasts, neurons, and blood vessel cells.Methods: We isolated anulus fibrosus cells from the nondegenerative intervertebral discs of adolescent (thirteen to sixteen-year-old) patients with idiopathic scoliosis and cultured the cells in vitro in induction media containing different stimuli. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Expression of markers of adipogenesis, osteogenesis, chondrogenesis, neurogenesis, and differentiation into endothelial lineages was determined with use of immunostaining, cytohistological staining, and reverse transcription-polymerase chain reaction.Results: Anulus fibrosus cells expressed several of the cell surface antigens that are sometimes associated with mesenchymal stem cells, including CD29, CD49e, CD51, CD73, CD90, CD105, CD166, CD184, and Stro-1, and two neuronal stem cell markers, nestin and neuron-specific enolase. Furthermore, varying the stimulants added to the induction media determined whether anulus fibrosus cells differentiated into adipocytes, osteoblasts, chondrocytes, neurons, or endothelial cells.Conclusions: Anulus fibrosus cells isolated from nondegenerative intervertebral discs can differentiate into adipocytes, osteoblasts, chondrocytes, neurons, and endothelial cells in vitro.