• 哈爾濱醫(yī)科大學(xué)附屬第二醫(yī)院肝膽胰腹腔鏡外科(哈爾濱150086);

目的 觀察凍存復(fù)蘇對(duì)人臍血(human umbilical cord blood)間充質(zhì)干細(xì)胞(mesenchymal stem cells, MSCs)生物學(xué)特性的影響.方法 體外分離、培養(yǎng)人臍血MSCs,傳代后,將第3代的MSCs加入含10%二甲基亞砜(DMSO)和90%胎牛血清的細(xì)胞凍存液中,
－196 ℃液氮保存4周,觀察比較凍存前及凍存復(fù)蘇后MSCs的形態(tài)、增殖及多向分化能力.結(jié)果 凍存前及凍存復(fù)蘇后,MSCs形態(tài)無(wú)明顯差別,均呈典型的梭形,MSCs貼壁生長(zhǎng); MSCs生長(zhǎng)曲線相似,凍存復(fù)蘇后的細(xì)胞生長(zhǎng)曲線略有下降,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P gt;0.05); MSCs經(jīng)脂肪誘導(dǎo)液誘導(dǎo)2周后,細(xì)胞漿中出現(xiàn)脂肪細(xì)胞所特有的脂肪滴,經(jīng)0.5% 油紅O染色,脂肪滴染為紅色,說(shuō)明MSCs有向脂肪細(xì)胞分化的能力; MSCs經(jīng)成骨誘導(dǎo)液誘導(dǎo)4周后,Von Kossa染色可見黑色的礦化結(jié)節(jié)沉積,鈣結(jié)節(jié)的形成為成骨細(xì)胞特有,說(shuō)明MSCs有向成骨細(xì)胞分化的能力.提示凍存復(fù)蘇后MSCs經(jīng)誘導(dǎo)仍然可以向脂肪細(xì)胞和成骨細(xì)胞分化,與凍存前無(wú)明顯差異.結(jié)論 人臍血MSCs經(jīng)凍存復(fù)蘇后,其生物學(xué)特性可獲良好保持.

引用本文： 吳德全,胡彥華,高峰,金光鑫. 凍存對(duì)人臍血間充質(zhì)干細(xì)胞生物學(xué)特性的影響. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008, 15(1): 36-40. doi: 復(fù)制

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2. Lee OK,Kuo TK,Chen WM,et al.Isolation of multipotent mesenchymal stem cells from umbilical cord blood[J].Blood,2004; 103(5)∶1669.
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20. 邢瑩,秦潔,曹孟德,等.細(xì)胞因子誘導(dǎo)人臍血間充質(zhì)干細(xì)胞分化過(guò)程中細(xì)胞巢蛋白、神經(jīng)絲亞單位和端粒酶逆轉(zhuǎn)錄酶mRNA的表達(dá)[J].鄭州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版),2005; 40(2)∶250.
21. 熊中華,許倩,陸德云,等.凍存人臍血間充質(zhì)干細(xì)胞向類肝細(xì)胞的誘導(dǎo)分化[J].中國(guó)組織工程研究與臨床康復(fù),2007; 11(7)∶1252.
22. 吳燕峰,包蓉,張緒超,等.人骨髓及臍血間充質(zhì)干細(xì)胞體外大量擴(kuò)增及建庫(kù)的質(zhì)量控制[J].實(shí)用醫(yī)學(xué)雜志,2006; 22(1)∶1.
  1. 1. Gang EJ,Hong SH,Jeong JA,et al.In vitro mesengenic potential of human umbilical cord blood-derived mesenchymal stem cells[J].Biochem Biophys Res Commun,2004; 321(1)∶102.
  2. 2. Lee OK,Kuo TK,Chen WM,et al.Isolation of multipotent mesenchymal stem cells from umbilical cord blood[J].Blood,2004; 103(5)∶1669.
  3. 3. Hong SH,Gang EJ,Jeong JA,et al.In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells[J].Biochem Biophys Res Commun,2005; 330(4)∶1153.
  4. 4. Gang EJ,Jeong JA,Hong SH,et al.Skeletal myogenic differentiation of mesenchymal stem cells isolated from human umbilical cord blood[J].Stem Cells,2004; 22(4)∶617.
  5. 5. Wild A,Jager M,Lensing-Hoehn S,et al.Growth behaviour of human mononuclear cells derived from bone marrow and cord blood on a collagen carrier for osteogenic regeneration[J].Biomed Tech (Berl),2004; 49(9)∶227.
  6. 6. 孫崇然,劉恩重.Von Kossa染色的方法改進(jìn)[J].哈爾濱醫(yī)科大學(xué)學(xué)報(bào),2006; 40(1)∶70.
  7. 7. 薛群,張學(xué)光,苗宗寧,等.胎盤間充質(zhì)干細(xì)胞樣細(xì)胞的培養(yǎng)及生物學(xué)特性的實(shí)驗(yàn)研究[J].蘇州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版),2005; 25(2)∶188.
  8. 8. Rosen ED,Walkey CJ,Puigserver P,et al.Transcriptional regulation of adipogenesis[J].Genes Dev,2000; 14(11)∶1293.
  9. 9. Yoshida S,Lshikawa F,Kawano N,et al.Human cord blood-derived cells generate insulin-producing cells in vivo[J].Stem Cell,2005; 23(9)∶1409.
  10. 10. 徐澤寬,苗毅.胰腺移植和胰島移植的現(xiàn)狀[J].中國(guó)普外基礎(chǔ)與臨床雜志,2003; 10(6)∶532.
  11. 11. 陳創(chuàng)奇,蘭平,詹文華.胰島移植與微循環(huán)的相關(guān)性研究[J].中國(guó)普外基礎(chǔ)與臨床雜志,2000; 7(6)∶411.
  12. 12. Rosada C,Justesen J,Melsvik D,et al.The human umbilical cord blood:a potential source for osteoblast progenitor cells[J].Calcif Tissue Int,2003; 72(2)∶135.
  13. 13. Yang SE,Ha CW,Jung M,et al.Mesenchymal stem/progenitor cells developed in cultures from UC blood[J].Cytotherapy,2004; 6(5)∶476.
  14. 14. Erices A,Conget P,Minguell JJ.Mesenchymal progenitor cells in human umbilical cord blood[J].Br J Haematol,2000; 109(1)∶235.
  15. 15. Romanov YA,Svintsitskaya VA,Smirnov VN.Searching for alternative sources of postnatal human mesenchymal stem cells:candidate MSC-like cells from umbilical cord[J].Stem Cells,2003; 21(1)∶105.
  16. 16. 張文元,楊亞冬,房國(guó)堅(jiān),等.不同凍存時(shí)間與溫度對(duì)兔骨髓基質(zhì)干細(xì)胞存活率的影響[J].中國(guó)臨床康復(fù),2006; 10(13)∶44.
  17. 17. 金光鑫,吳德全.臍帶血間充質(zhì)干細(xì)胞研究進(jìn)展[J].國(guó)際移植與血液凈化雜志,2006; 4(5)∶45.
  18. 18. Feldmann RE Jr,Bieback K,Maurer MH,et al.Stem cell proteomes:a profile of human mesenchymal stem cells derived from umbilical cord blood[J].Electrophoresis,2005; 26(14)∶2749.
  19. 19. 侯玲玲,曹華,魏國(guó)榮,等.人臍血間充質(zhì)干細(xì)胞體外擴(kuò)增和向神經(jīng)元樣細(xì)胞定向誘導(dǎo)分化的研究[J].中華血液學(xué)雜志,2002; 23(8)∶415.
  20. 20. 邢瑩,秦潔,曹孟德,等.細(xì)胞因子誘導(dǎo)人臍血間充質(zhì)干細(xì)胞分化過(guò)程中細(xì)胞巢蛋白、神經(jīng)絲亞單位和端粒酶逆轉(zhuǎn)錄酶mRNA的表達(dá)[J].鄭州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版),2005; 40(2)∶250.
  21. 21. 熊中華,許倩,陸德云,等.凍存人臍血間充質(zhì)干細(xì)胞向類肝細(xì)胞的誘導(dǎo)分化[J].中國(guó)組織工程研究與臨床康復(fù),2007; 11(7)∶1252.
  22. 22. 吳燕峰,包蓉,張緒超,等.人骨髓及臍血間充質(zhì)干細(xì)胞體外大量擴(kuò)增及建庫(kù)的質(zhì)量控制[J].實(shí)用醫(yī)學(xué)雜志,2006; 22(1)∶1.