Isolation and characterization of side population stem cells in articular synovial tissue.
Isolation and characterization of side population stem cells in articular synovial tissue.
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DOI：
10.1186/1471-2474-9-86
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发表时间：
2008-06-12
影响因子：
2.3
通讯作者：
Hamanishi C
中科院分区：
文献类型：
--
作者：
Teramura T;Fukuda K;Kurashimo S;Hosoi Y;Miki Y;Asada S;Hamanishi C
自体软骨细胞植入是一种成熟的修复退行性关节软骨的技术。近年来，检测具有强烈外排Hoechst 33342 （Ho）荧光染料能力的侧群（SP）细胞作为一种干细胞分离方法备受关注。虽然滑膜组织的SP细胞被认为是组织工程的良好来源，但它们在滑膜组织中的精确特性尚未确定。牛掌指关节滑膜组织被用作干细胞来源。为了有效地收集干细胞，我们首先在含有可变浓度FBS的培养基中制备预培养物，然后分选4天。使用细胞分选器和Ho-dye，然后分离出富含干细胞的低染色群体。为了确定干细胞的特征，采用实时荧光定量PCR技术对CD34、Flk-1、c-Kit、Abcg-2等特异性标记基因进行鉴定。将分选的SP细胞培养于添加bFGF、SCF和纤维连接蛋白的干细胞培养基中，评估其向软骨细胞、骨细胞和肌细胞分化的潜力。在含1%FBS的培养基中预培养，滑膜组织SP细胞从占总细胞群的2%增加到约占总细胞群的10%。分选后的SP细胞表达CD34、Flk-1、c-Kit、Abcg-2和Mdr-1 -都是干细胞特性的重要标记基因。SP细胞可在体外进一步扩增，同时保持标志物基因表达、Ho-dye外排电位和向软骨细胞、骨细胞和肌细胞的多向分化等干细胞潜能。在本研究中，我们证明了从牛滑膜中有效地收集具有突出干细胞特性的细胞作为SP部分。此外，我们还描述了一种有效的分离方法和体外扩增的培养条件，以保持其重要特征。我们的结果表明滑膜组织的SP细胞可能是细胞移植的重要候选来源。
Autologous chondrocyte implantation is an established technique for the repair of degenerated articular cartilage. Recently, the detection of side population (SP) cells, which have the ability to strongly efflux Hoechst 33342 (Ho) fluorescence dye, has attracted attention as a method of stem cell isolation. Although SP cells from synovial tissue were expected to be an excellent source for this tissue engineering, their precise character in the synovial tissue has not been determined. Synovial tissues from bovine metacarpophalangeal joints were used as a stem cell source. For efficient collection of stem cells, we first prepared a preculture before sorting in medium containing FBS at variable concentrations for 4 days. Using a cell sorter and the Ho-dye, a poorly stained population enriched with stem cells was then isolated. To determine the characteristics of the stem cells, specific marker genes such as CD34, Flk-1, c-Kit, Abcg-2 were identified by real-time PCR. Sorted SP cells were cultured in a stem cell medium supplemented with bFGF, SCF and fibronectin, and evaluated for their differentiation potentials into chondrocytes, osteocytes and myocytes. SP cells of synovium tissue were increased from 2% of the total cell population to approximately 10% of the total cells by preculture in the 1%FBS contained medium. Sorted SP cells expressed CD34, Flk-1, c-Kit, Abcg-2 and Mdr-1 -all are important marker genes for stem cell characteristics. The SP cells could be further expanded ex vivo while maintaining stem cell potentials such as marker gene expression, Ho-dye efflux potential and multiple differentiation potentials into chondrocyte, osteocyte and myocyte. In the present study, we demonstrated that the cells with outstanding stem cell properties were efficiently collected as a SP fraction from bovine synovial membrane. Furthermore, we have described an efficient isolation method and the culture conditions for ex vivo expansion that maintains their important characteristics. Our results suggest that the SP cells of synovium tissue might be important candidates as sources for cell transplantation.
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影响因子：
13.3
作者：
Mauduit, C;Hamamah, S;Benahmed, M
通讯作者：
Benahmed, M
影响因子：
5.2
作者：
Levenstein, Mark E.;Ludwig, Tenneille E.;Thomson, James A.
通讯作者：
Thomson, James A.
影响因子：
20.3
作者：
KELLER, JR;ORTIZ, M;RUSCETTI, FW
通讯作者：
RUSCETTI, FW
影响因子：
20.3
作者：
Dao, MA;Hashino, K;Nolta, JA
通讯作者：
Nolta, JA
影响因子：
4.6
作者：
Lassalle, B;Bastos, H;Allemand, I
通讯作者：
Allemand, I