[1]黄团结),马珊珊),邢 衢),等.rhMG53作为低温保护剂对小鼠BMSCs生物学特性的影响[J].郑州大学学报(医学版),2018,(06):710-714.[doi:10.13705/j.issn.1671-6825.2018.04.095]
 HUANG Tuanjie),MA Shanshan),XING Qu),et al.Effects of rhMG53 as a special cryoprotectant on biological characteristics of mouse bone marrow mesenchymal stem cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2018,(06):710-714.[doi:10.13705/j.issn.1671-6825.2018.04.095]
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rhMG53作为低温保护剂对小鼠BMSCs生物学特性的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2018年06期
页码:
710-714
栏目:
论著
出版日期:
2018-11-20

文章信息/Info

Title:
Effects of rhMG53 as a special cryoprotectant on biological characteristics of mouse bone marrow mesenchymal stem cells
作者:
黄团结1)马珊珊1)邢 衢1)周建康1)刘腾飞1)周馨魁1)王亚苹1)杨 波2)麻建杰3)关方霞12)
1)郑州大学生命科学学院 郑州450001;2)郑州大学第一附属医院神经外科 郑州450052;3)俄亥俄州立大学Davis心肺研究所外科部 美国俄亥俄州哥伦布 43210
Author(s):
HUANG Tuanjie1) MA Shanshan1) XING Qu1) ZHOU Jiankang1) LIU Tengfei1) ZHOU Xinkui1) WANG Yaping1) YANG Bo2) MA Jianjie3) GUAN Fangxia12)
1)School of Life Sciences,Zhengzhou University,Zhengzhou 450001;2)Department of Neurosurgery,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;3)Department of Surgery,Davis Heart and Lung Research Institute,The Ohio State University,Columbus,OH 43210 USA
关键词:
rhMG53 骨髓间充质干细胞 冷冻保存 保护剂 生物学特性
Keywords:
rhMG53 bone marrow mesenchymal stem cell cryopreservation protective agent biological characteristics
分类号:
Q28
DOI:
10.13705/j.issn.1671-6825.2018.04.095
摘要:
目的:探讨重组人MG53蛋白(rhMG53)作为干细胞低温保护剂的特殊组分对小鼠骨髓间充质干细胞(BMSCs)冷冻保存后生物学特性的影响。方法:常规培养BMSCs,分为对照组(DMEM/F12培养基、FBS、DMSO的体积比为5:4:1并添加30 mg/L BSA)和rhMG53组(DMEM/F12培养基、FBS、DMSO的体积比为5:4:1并添加30 mg/L rhMG53)冻存。冻存1 a后,复苏细胞,qRT-PCR检测两组细胞冻存复苏后干性相关基因Nanog、OCT4和SOX2 mRNA的表达; CCK-8法检测细胞增殖; 台盼蓝染色检测细胞存活能力; 茜素红S染色和油红O染色分别检测两组干细胞的成骨和成脂分化; 免疫荧光检测两组细胞成神经分化相关因子DCX和NSE的表达。结果:与对照组相比,rhMG53组BMSCs的Nanog、OCT4和SOX2 mRNA表达无明显变化(P>0.05),但细胞增殖能力增强,存活率升高,成骨、成脂和成神经分化效率增加(P均<0.05)。结论:rhMG53作为保护剂的特殊组分对冷冻保存的BMSCs有较好的保护效果。
Abstract:
Aim:To observe the effects of recombinant human MG53 protein(rhMG53),as a special component of the low temperature protective agent of stem cells, on the biological characteristics of mouse bone marrow mesenchymal stem cells(BMSCs).Methods:BMSCs were expanded and defined as control group(VDMEM/F12:VFBS:VDMSO=5:4:1,added 30 mg/L BSA)and rhMG53 group(VDMEM/F12:VFBS:VDMSO=5:4:1, added 30 mg/L rhMG53).After one year of cryopreservation,BMSCs were resuscitated.The expressions of stemness related genes Nanog,OCT4 and SOX2 were detected by qRT-PCR; CCK-8 was used to detect cell proliferation; Trypan blue staining was used to detect cell viability.Alizarin red S staining and oil red O staining were used to detect the osteogenic and adipogenic differentiation of the stem cells.Immunofluorescence was used to detect the expressions of DCX and NSE genes related to neural differentiation.Results: Compared with control group,there were no differences in the expressions of the stemness related genes in the rhMG53 group(P>0.05),while the proliferation ability of BMSCs was enhanced,the cells survival rate was increased,and the efficiency of osteogenesis, adipogenesis and neurogenic differentiation was promoted(P<0.05).Conclusion:rhMG53,as a specific components of low temperature protective agent,does not affect the stemness of BMSCs,and could improve the proliferation and survival abilities,as well as osteogenic,adipogenic,and neural differentiation efficiency of the stem cells after cryopreservation and resuscitation.

参考文献/References:

[1] DERUBEIS AR,CANCEDDA R.Bone marrow stromal cells(BMSCs)in bone engineering: limitations and recent advances[J].Ann Biomed Eng,2004,32(1):160
[2] CAPLAN AI.MSCs:the sentinel and safe-guards of injury[J].J Cell Physiol,2016,231(7):1413
[3] ZHANG J,HUANG XW,WANG HJ,et al.The challenges and promises of allogeneic mesenchymal stem cells for use as a cell-based therapy[J].Stem Cell Res Ther,2015,6(6):1
[4] WOODS E,THIRUMALA SJ,BADHE-BUCHANAN SS,et al.Off the shelf cellular therapeutics:factors to consider during cryopreservation and storage of human cells for clinical use[J].Cytotherapy,2016,18(6):697
[5] MARQUEZ-CURTIS LA,JANOWSKA-WIECZOREK A,MCGANN LE,et al.Mesenchymal stromal cells derived from various tissues:biological, clinical and cryopreservation aspects[J].Cryobiology,2015,71(2):181
[6] YONG KW,PINGGUAN-MURPHY B,XU F,et al.Phenotypic and functional characterization of long-term cryopreserved human adipose-derived stem cells[J].Sci Rep,2015,5:9596
[7] BEIER AF,SCHULZ JC,DÖRR D,et al.Effective surface-based cryopreservation of human embryonic stem cells by vitrification[J].Cryobiology,2011,63(3):175
[8] MIRANDA-SAYAGO JM,FERNANDEZ-ARCAS N,BENITO C,et al.Evaluation of a low cost cryopreservation system on the biology of human amniotic fluid-derived mesenchymal stromal cells[J].Cryobiology,2012,64(3):160
[9] 关方霞,马珊珊,黄团结,等. 一种包含rhMG53的人脐带间充质干细胞低温保护剂:CN107156107A[P]. 2017-09-15.
[10]CAI C,MASUMIYA H,WEISLEDER N,et al.MG53 nucleates assembly of cell membrane repair machinery[J].Nat Cell Biol,2009,11(1):56
[11]WANG X,XIE W,ZHANG Y,et al.Cardioprotection of ischemia/reperfusion injury by cholesterol-dependent MG53-mediated membrane repair[J].Circ Res,2010,107(1):76
[12]CAI CX,LIN PH,ZHU H,et al.Zinc binding to MG53 protein facilitates repair of injury to cell membranes[J].J Biol Chem,2015,290(22):13830
[13]WEISLEDER N,TAKIZAWA N,LIN P,et al.Recombinant MG53 protein modulates therapeutic cell membrane repair in treatment of muscular dystrophy[J].Sci Transl Med,2012,4(139):139ra85
[14]邢衢,马珊珊,王欣欣,等.人脐带间充质干细胞神经分化微环境的优化[J].郑州大学学报(医学版),2016,51(4):469
[15]黄团结,宋及时,马珊珊,等.重组人MG53蛋白对人脐带间充质干细胞氧化损伤的保护作用[J].中华细胞与干细胞杂志:电子版,2017,7(1):35
[16]LIU J,ZHU H,ZHENG Y,et al.Cardioprotection of recombinant human MG53 protein in a porcine model of ischemia and reperfusion injury[J].J Mol Cell Cardiol,2015,80(3):10
[17]YAO YG,ZHANG B,ZHU H,et al.MG53 permeates through blood-brain barrier to protect ischemic brain injury[J].Oncotarget,2016,7(16):22474

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备注/Memo

备注/Memo:
【基金项目】国家自然科学基金资助项目(81601078,81471306); 河南省高校科技创新团队项目(15IRTSTHN022); 河南省科技创新人才计划项目(154200510008); 河南省国际人才合作项目(2016GH03); 河南省重点科技攻关项目(152102310272)
【作者简介】关方霞,通信作者,女,1969年2月生,博士,教授,研究方向:干细胞与再生医学,E-mail:guanfangxia@126.com; 马珊珊,通信作者,女,1984年5月生,博士,副教授,研究方向:干细胞与神经功能修复,E-mail:mashanshan84@163.com
更新日期/Last Update: 2018-11-20