人外周血单个核细胞与脐带间充质干细胞共培养的AFM研究

doi:Q819

中国生物工程杂志

2010, Vol. 30

Issue (08): 6-12 DOI: Q819

研究报告

人外周血单个核细胞与脐带间充质干细胞共培养的AFM研究

龚超群¹,蔡继业^1**,郜世隽²,王秋兰¹,金花¹

1.暨南大学生命科学技术学院化学系广州 510632
2.暨南大学医学院微生物学免疫学教研室广州 510632

The Process of Coculturing Peripheral Blood Mononuclear Cells and Human Umbilical Cord Derived Mesenchymal Stem Cells Observed with Atomic Force Microscope

GONG Chao-qun¹,CAI Ji-ye¹,GAO Shi-jun²,WANG Qiu-lan¹,JIN Hua¹

1.Department of Chemistry, College of Life Science and Technology , Jinan University, Guangzhou 510632,China
2.Department of Microbiology and Immunology, Medical College, Jinan University, Guangzhou 510632, China

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摘要：

采用原子力显微镜与倒置显微镜在细胞层次上观察了人外周单个核细胞 (PBMCs)与同种异源脐带间充质干细胞(hUC-MSCs)共培养的过程，并在单细胞水平上分析了共培养前后人外周单个核细胞的形貌和生物物理性质。结果发现：共培养后贴壁人外周单个核细胞的形态发生了很大的改变，并且表面分布着大小不一的颗粒状聚合物。利用 AFM高空间分辨的力位移曲线测量系统，发现共培养72 h后培养上清中人外周单个核细胞、贴壁的人外周单个核细胞的粘滞力分别是单纯培养72 h的人外周单个核细胞的2倍、5倍，而细胞的硬度分别是单纯培养人外周单个核细胞的1.5倍、2倍。CCK-8检测提示，共培养过程中，干细胞的生长与外周血单个核细胞的生长出现了竞争作用。通过AFM探测人外周单个核细胞与脐带间充质干细胞共培养的可视化数据，有助于更好地了解间充质干细胞与外周血单个核细胞的相互作用。

关键词： 脐带间充质干细胞; 外周血单个核细胞; 原子力显微镜; 倒置荧光显微镜

Abstract:

The inverted microscope was used to observe the process that peripheral blood mononuclearcells (PBMCs) were cocultured with allogeneic human umbilical cord derived mesenchymal stem cells ( hUC- MSCs). The atomic force microscope was used to analyze the morphology and mechanical property after two kinds of cells were cocultured. The result showed that the morphology of adherence PBMCs changed greatly after cocultured, and was coated with different grain sizes of polymers. From the high spatial resolution of AFM force-curves, the adhesion force and stiffness values of PBMCs cocultured with hUC-MSCs were stronger than those of controlled PBMCs. CCK-8 indicated that PBMCs may appear to compete with hUC- MSCs in the co-culture system. The visualized data obtained from AFM can provide a better understanding the interactive mechanism of hUC-MSCs and PBMCs.

Key words: Human umbilical cord mesenchymal stem cells Peripheral blood mononuclear cells Atomic force microscope (AFM) Inverted microscopemicroscope

收稿日期: 2009-11-16 出版日期: 2010-08-25

基金资助:

海外及港澳学者合作研究基金(30828028)、国家自然科学基金(30872404) 资助项目

通讯作者: 蔡继业 E-mail: tjycai@jnu.edu.cn

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引用本文:

龚超群蔡继业郜世隽王秋兰金花. 人外周血单个核细胞与脐带间充质干细胞共培养的AFM研究[J]. 中国生物工程杂志, 2010, 30(08): 6-12.

GONG Chao-Qun, CA Ji-Ye, GAO Shi-Juan, WANG Qiu-Lan, JIN Hua. The Process of Coculturing Peripheral Blood Mononuclear Cells and Human Umbilical Cord Derived Mesenchymal Stem Cells Observed with Atomic Force Microscope. China Biotechnology, 2010, 30(08): 6-12.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/Q819 或 https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I08/6

［1］ Koc O N, Day J, Nieder M, et al. Allogeneic mesenchymal stem cell infusion for treatment of metachromatic leukodystrophy (MLD and Hurler syndrome (MPSI H). Bone Marrow Transplant, 2002, 30 (4): 215222.
［2］ Chen S L, Fang W, Ye F, et al. Effect on left ventricular function of in tracoronary transplantation of autologous bone marrow mesenchymal stem cell in patients with acute myocardial infarction. American Journal Cardiology, 2004, 94 (1): 9295.
［3］王茜，杨琨，白海,等. 人脐血间充质干细胞抑制异体 T外周血单个核细胞反应的实验及临床意义. 第四军医大学学报，2007, 28 (18)：16491651. Wang Q, Yang K, Bai H, et al. J Fourth Mil Med Univ, 2007, 28 (18)：16491651.
［4］白海，王茜，吴涛，等. 人脐血间充质干细胞培养及对异体外周血T外周血单个核细胞增殖的影响. 中国组织工程研究与临床康复，2008，12(3)：338441. Bai H, Wang Q, Wu T, et al. Journal of Clinical Rehabilitative Tissue Engineering Research，2008，12(3)：338441.
［5］王蒙，杨媛，杨东明，等. 人脐血源性 MSCs的免疫调节作用. 免疫学杂志，2007, 23(3)：287294. Wang M, Yang Y, Yang D M, et al. Immunological Journal, 2007, 23(3):287294.
［6］陶艳玲, 张颢. 脐血间充质干细胞对 T外周血单个核细胞增殖和激活影响的实验研究.中国实用儿科杂志, 2007, 23(3): 287294. Tao Y L, Zhang H. Chinese Journal of Practical Pediatrics, 2007, 23(3):287294.
［7］ Wonil O, DalSoo K, Yoon S Y, et al. Immunological properties of umbilical cord bloodderived mesenchymal stromal cells. Cellular Immunology, 2008，251：116123.
［8］ Mark L W, Cameron A, Satish M, et al. Immune properties of human umbilical cord Wharton’s Jellyderived cells. Stem Cells，2008，26：28652874.
［9］ Evans E A, Calderwood D A. Forces and bond dynamics in cell adhesion. Science, 2007, 316 (5828) : 11481153.
［10］ Wu Z Z，Zhang G, Long M, et al.Comparison of the viscoelastic properties of normal hepatocytes and hepato cellular carcinoma cells under cytoskeletal perturbation. Biorheology, 2000, 37 (4)：279290.
［11］ Cross S E, Jin Y S, Rao J Y,et al. Nanomechanical analysis of cells from cancer patients. Nature Nanotechnology, 2007, 2(12): 780783.
［12］ Crick S L, Yin F C P. Assessing micromechanical properties of cells with atomic force microscopy: importance of the contact point. Biomechanics and Modeling in Mechanobiology, 2007, 6(3): 199210.
［13］ Suresh S. Biomechanics and biophysics of cancer cell. Acta Biomaterialia, 2007, 3(4): 413438.
［14］ Li Q S, Lee G Y H, Ong C N, et al. AFM indentation study of breast cancer cells. Biochemical and Biophysical Research Communication, 2008, 374(4):609613.
［15］ Hsieh C H, Lin Y H, Lin S, et al. Surface ultrastructure and mechanical property of human chondrocyte revealed by atomic force microscopy. Osteoarthritis and Cartilage, 2008, 16(4): 480488.
［16］ Karahuseyinoglu Cinar Kilic. Biology of stem cells human umbilical cord stroma. In Situ and In Vitro Survey Stem Cells, 2007, 25: 319331.
［17］ Weiss M L, Medicetty S, Bledsoe A R, et al. Human umbilic cord matrix stem cells: preliminary characterization and effect transplantation in a rodent model of Parkinson’s disease. Stem Cells, 2006, 24: 781792.
［18］ Conconi M T, Burra P, DiLiddo R, et al . CD105 (+) cells from Wharton’s jelly show in vitro and in vivo myogenic differentative potential. Int J Mol Med, 2006, 18 (6) : 10891096.
［19］ Lupatov A Y, Karalkin P A, Suzdal’tseva Y G, et al. Cytofluorometric analysis of phenotypes of human bone marrow and umbilcal fibroblastlike cells. Bull Exp Biol Med, 2006, 142 (4)：521526.

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