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[1]王蕭楓,童培建,金紅婷,等.兔骨髓間充質(zhì)干細(xì)胞的體外培養(yǎng)及相關(guān)檢測(cè)的實(shí)驗(yàn)研究[J].中醫(yī)正骨,2011,23(11):3-6.
　WANG Xiao-feng*,TONG Pei-Jian,JIN Hong-ting,et al.Approaches To Isolate,Culture and Identify The Rabbit Marrow Mesenchymal Stem Cells[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2011,23(11):3-6.
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兔骨髓間充質(zhì)干細(xì)胞的體外培養(yǎng)及相關(guān)檢測(cè)的實(shí)驗(yàn)研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第23卷
期數(shù):
2011年11期
頁(yè)碼:
3-6
欄目:
基礎(chǔ)研究
出版日期:
2011-11-30

文章信息/Info

Title:
Approaches To Isolate,Culture and Identify The Rabbit Marrow Mesenchymal Stem Cells
作者:
王蕭楓1; 童培建2; 金紅婷2; 吳銀生1
1.浙江省溫州市中西醫(yī)結(jié)合醫(yī)院,浙江 溫州 325000; 2.浙江中醫(yī)藥大學(xué),浙江 杭州 310053
Author(s):
WANG Xiao-feng*; TONG Pei-Jian; JIN Hong-ting; et al.*
Hospital of the Integration of Traditional Chinese and Western Medicine of Wenzhou City,Wenzhou 325000,Zhejiang,China
關(guān)鍵詞:
間充質(zhì)干細(xì)胞 體外培養(yǎng) 誘導(dǎo)分化 流式細(xì)胞術(shù)
Keywords:
Mesenchymal stem cells;  Cell culture techniques
摘要:
目的:建立體外培養(yǎng)擴(kuò)增、檢測(cè)鑒定兔骨髓間充質(zhì)干細(xì)胞的方法,觀察其生物學(xué)特性。方法:采用密度梯度離心法和貼壁分離法相結(jié)合,體外培養(yǎng)擴(kuò)增兔骨髓間充質(zhì)干細(xì)胞,觀察細(xì)胞生長(zhǎng)速度和形態(tài)變化; 選擇第3代細(xì)胞分別以特定培養(yǎng)液誘導(dǎo),觀察其向成骨細(xì)胞、成脂細(xì)胞、成軟骨細(xì)胞的分化能力,采用流式細(xì)胞學(xué)方法分別檢測(cè)CD29、CD44、CD34、CD45的表達(dá)情況。結(jié)果:培養(yǎng)的細(xì)胞長(zhǎng)梭形,呈成纖維細(xì)胞樣生長(zhǎng),具有塑料貼附性、傳代細(xì)胞生長(zhǎng)迅速,在誘導(dǎo)下均能分化為成熟的成骨細(xì)胞、軟骨細(xì)胞和脂肪細(xì)胞。流式結(jié)果表明,CD29陽(yáng)性細(xì)胞比率92.5%,CD44陽(yáng)性細(xì)胞比率86%; CD34陽(yáng)性比率0.77%,CD45陽(yáng)性細(xì)胞比率0.32%。結(jié)論:應(yīng)用密度梯度分離法和貼壁篩選法相結(jié)合能從兔骨髓中分離培養(yǎng)出高純度的兔MSCs,生長(zhǎng)旺盛、增殖能力強(qiáng),在條件誘導(dǎo)下具有向成骨細(xì)胞、脂肪細(xì)胞和軟骨細(xì)胞分化的潛能。
Abstract:
Objective:To explore the approaches to isolate,culture and identify the bone marrow mesenchymal stem cells(MSCs)of rabbits in vitro.Methods:Rabbit MSCs were extracted through the density gradient centrifugation and separation of the combination adherent,cell growth rate and morphological changes was observed using the inverted microscope,select the P3 cells induced using special media supplemented,observe the feasibility of differentiation into bone cells,fat cells,cartilage cells.Expression of CD29,CD44,CD34,CD45 was detected using flow cytometry.Results:The cultured cells showing long fusiform and fibroblast-like growth,plastic attachment and rapid growth of passage cell,they can differentiate into mature bone cells,cartilage cells and fat cells under induction.Flow cytometry results showed that for the P3 cells,about 92.5% of the cells expressed CD29,about 86% of the cells expressed CD44,0.77% of the cells expressed CD34,0.32% of the cells expressed CD45.Conclusion:The density gradient separation and screening adherence combination isolated from rabbit bone marrow in vitro with high purity MSCs,vigorous growth,proliferation ability,under the conditions with the induction of osteoblasts,adipose cells and cartilage cells potential.

參考文獻(xiàn)/References:

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更新日期/Last Update: 1900-01-01