Effect of Cryopreservation and Resuscitation on Biological Characteristics of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

WU Dequan , HU Yanhua , GAO Feng , JIN Guangx in

Department of Hepatobiliary pancreatic and Laparoscopic Surgery , The Second Affiliated Hospital of Harbin Medical Universit y , Harbin 150086 , Chi na;

Corresponding author：

Keywords：

Mesenchymal stem cells derived f rom human umbilical cord blood, 　Cryopreservation, 　Biological characteristics

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Objective 　To observe the effects of cryopreservation and resuscitation on the biological characteristics of mesenchymal stem cells (MSCs) derived f rom human umbilical cord blood. Methods 　MSCs were isolated and cultured f rom human umbilical cord blood in vitro. The cells were passaged , and the third generation of MSCs were cryopreserved in－196 ℃ liquid nitrogen for 4 weeks with cryopreservation medium , which contained 10 % dimethyl sulfoxide (DMSO) and 90 % fetal calf serum ( FCS) . The morphology , proliferation and differentiation of MSCs were investigated and compared with those of MSCs before cryopreservation. Results 　There was no significant difference of morphology between pre-cryopreserved MSCs and the ones af ter resuscitation. It was observed that all MSCs were spindle-shaped and showed adherence growth characteristic before and af ter cryopreservation. The cell growth curves of MSCs were also similar before and af ter cryopreservation. Even though the curve of resuscitated MSCs descended a little as compared with that of pre-cryopreserved MSCs , there was no significant difference ( P gt; 0. 05) . After 2-week adipocytic differentiation induction , fat drops could be found in the kytoplasm of MSCs and they were red when stained with oil-red O staining , which suggested that MSCs could be induced and differentiated into adipocytes. Af ter 4-week osteoblastic differentiation induction , MSCs could be induced and differentiated into osteoblasts , and calcium node showed black when stained with Von Kossa staining. There were no significant changes of the differentiating ability of MSCs into adipocyte and osteoblast before and after cryopreservation. Conclusion 　MSCs derived from human umbilical cord blood maintains their biological characteristics af ter cryopreservation and resuscitation.

Citation： WU Dequan,HU Yanhua,GAO Feng,JIN Guangx in. Effect of Cryopreservation and Resuscitation on Biological Characteristics of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2008, 15(1): 36-40. doi: Copy

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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. 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. 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. 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. 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. 孙崇然,刘恩重.Von Kossa染色的方法改进[J].哈尔滨医科大学学报,2006; 40(1)∶70.
7. 薛群,张学光,苗宗宁,等.胎盘间充质干细胞样细胞的培养及生物学特性的实验研究[J].苏州大学学报(医学版),2005; 25(2)∶188.
8. Rosen ED,Walkey CJ,Puigserver P,et al.Transcriptional regulation of adipogenesis[J].Genes Dev,2000; 14(11)∶1293.
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. 徐泽宽,苗毅.胰腺移植和胰岛移植的现状[J].中国普外基础与临床杂志,2003; 10(6)∶532.
11. 陈创奇,兰平,詹文华.胰岛移植与微循环的相关性研究[J].中国普外基础与临床杂志,2000; 7(6)∶411.
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. 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. Erices A,Conget P,Minguell JJ.Mesenchymal progenitor cells in human umbilical cord blood[J].Br J Haematol,2000; 109(1)∶235.
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. 张文元,杨亚冬,房国坚,等.不同冻存时间与温度对兔骨髓基质干细胞存活率的影响[J].中国临床康复,2006; 10(13)∶44.
17. 金光鑫,吴德全.脐带血间充质干细胞研究进展[J].国际移植与血液净化杂志,2006; 4(5)∶45.
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. 侯玲玲,曹华,魏国荣,等.人脐血间充质干细胞体外扩增和向神经元样细胞定向诱导分化的研究[J].中华血液学杂志,2002; 23(8)∶415.
20. 邢莹,秦洁,曹孟德,等.细胞因子诱导人脐血间充质干细胞分化过程中细胞巢蛋白、神经丝亚单位和端粒酶逆转录酶mRNA的表达[J].郑州大学学报(医学版),2005; 40(2)∶250.
21. 熊中华,许倩,陆德云,等.冻存人脐血间充质干细胞向类肝细胞的诱导分化[J].中国组织工程研究与临床康复,2007; 11(7)∶1252.
22. 吴燕峰,包蓉,张绪超,等.人骨髓及脐血间充质干细胞体外大量扩增及建库的质量控制[J].实用医学杂志,2006; 22(1)∶1.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of Cryopreservation and Resuscitation on Biological Characteristics of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

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