Differential expression of CCN-family members in primary human bone marrow-derived mesenchymal stem cells during osteogenic, chondrogenic and adipogenic differentiation.
Differential expression of CCN-family members in primary human bone marrow-derived mesenchymal stem cells during osteogenic, chondrogenic and adipogenic differentiation.
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在成骨，软骨源性和脂肪生成分化期间，在原发性人骨髓衍生的间充质干细胞中，CCN家庭成员的差异表达。
DOI：
10.1186/1478-811x-3-5
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发表时间：
2005-03-17
影响因子：
8.4
通讯作者：
Jakob, Franz
中科院分区：
文献类型：
--
作者：
Schutze, Norbert;Noth, Ulrich;Schneidereit, Jutta;Hendrich, Christian;Jakob, Franz
人富含半胱氨酸蛋白61（CYR 61，CCN 1）以及CCN基因家族的其他成员在细胞过程如增殖、粘附、迁移和存活中起重要作用。这些细胞事件在骨重建中进行的复杂细胞相互作用中特别重要。以前，我们分析了CYR 61/CCN 1作为细胞外信号分子在人成骨细胞中的作用。由于骨髓间充质干细胞是骨中多种分化途径的重要祖细胞，并且具有越来越大的再生医学潜力，因此，本文旨在分析CCN家族成员在骨髓来源的人间充质干细胞中的表达以及沿着成骨、成脂和成软骨分化。从接受全髋关节置换术的患者的股骨头获得人间充质干细胞的原代培养物。在单层培养中分化成脂肪细胞和成骨细胞，在高密度细胞团培养中诱导分化成软骨细胞。对于任一途径，通过RT-PCR在mRNA水平分析已建立的分化标志物和CCN成员，并通过免疫细胞化学分析CYR 61/CCN 1蛋白。RT-PCR和组织化学分析显示了分化细胞的适当表型（茜素红S、油红O、阿尔新蓝、碱性磷酸酶;骨钙素、I、II、IX、X型胶原、cbfa 1、PPARγ、聚集蛋白聚糖）。间充质干细胞表达CYR 61/CCN 1、CTGF/CCN 2、CTGF-L/WISP 2/CCN 5和WISP 3/CCN 6。CYR 61/CCN 1在成骨、成脂和成软骨分化过程中表达明显下降。免疫细胞化学分析证实了这些结果。WISP 2/CCN 5 RNA表达在成脂分化过程中下降，WISP 3/CCN 6 RNA表达在成软骨分化过程中显著降低。在间充质干细胞分化为成骨细胞、脂肪细胞和软骨细胞的过程中，CYR 61/CCN 1表达的降低表明CYR 61/CCN 1在维持干细胞表型中的特定作用。CTGF/CCN 2、WISP 2/CCN 5、WISP 3/CCN 6和主要是CYR 61/CCN 1的差异表达表明，CCN家族的这些成员可能是骨髓间充质干细胞在调节增殖和启动特异性分化途径中的重要调节剂。
The human cysteine rich protein 61 (CYR61, CCN1) as well as the other members of the CCN family of genes play important roles in cellular processes such as proliferation, adhesion, migration and survival. These cellular events are of special importance within the complex cellular interactions ongoing in bone remodeling. Previously, we analyzed the role of CYR61/CCN1 as an extracellular signaling molecule in human osteoblasts. Since mesenchymal stem cells of bone marrow are important progenitors for various differentiation pathways in bone and possess increasing potential for regenerative medicine, here we aimed to analyze the expression of CCN family members in bone marrow-derived human mesenchymal stem cells and along the osteogenic, the adipogenic and the chondrogenic differentiation. Primary cultures of human mesenchymal stem cells were obtained from the femoral head of patients undergoing total hip arthroplasty. Differentiation into adipocytes and osteoblasts was done in monolayer culture, differentiation into chondrocytes was induced in high density cell pellet cultures. For either pathway, established differentiation markers and CCN-members were analyzed at the mRNA level by RT-PCR and the CYR61/CCN1 protein was analyzed by immunocytochemistry. RT-PCR and histochemical analysis revealed the appropriate phenotype of differentiated cells (Alizarin-red S, Oil Red O, Alcian blue, alkaline phosphatase; osteocalcin, collagen types I, II, IX, X, cbfa1, PPARγ, aggrecan). Mesenchymal stem cells expressed CYR61/CCN1, CTGF/CCN2, CTGF-L/WISP2/CCN5 and WISP3/CCN6. The CYR61/CCN1 expression decreased markedly during osteogenic differentiation, adipogenic differentiation and chondrogenic differentiation. These results were confirmed by immuncytochemical analyses. WISP2/CCN5 RNA expression declined during adipogenic differentiation and WISP3/CCN6 RNA expression was markedly reduced in chondrogenic differentiation. The decrease in CYR61/CCN1 expression during the differentiation pathways of mesenchymal stem cells into osteoblasts, adipocytes and chondrocytes suggests a specific role of CYR61/CCN1 for maintenance of the stem cell phenotype. The differential expression of CTGF/CCN2, WISP2/CCN5, WISP3/CCN6 and mainly CYR61/CCN1 indicates, that these members of the CCN-family might be important regulators for bone marrow-derived mesenchymal stem cells in the regulation of proliferation and initiation of specific differentiation pathways.