猪胎盘绒毛滋养层细胞体外原代培养

doi:10.11843/j.issn.0366-6964.2017.01.009

Abstract

Abstract:

This research was conducted to establish a simple and effective method for isolation and purification of pig chorionic trophoblast cells. Single cell suspension was prepared from pig fullterm placentas by the method of trypsin-DNaseⅠdigestion and two density (35% and 45%, v/v) Percoll gradient centrifugation.Morphology observation,growth curve measurement, immunofluorescence and transmission electron microscopy (TEM) were applied to cells identification. The results showed that the cells were polygon or quasi-circular in shape, which exhibited epithelioid and paving-like spreading growth. After 48 hours culture, some cells started developing into syncytiotrophoblasts. Immunofluorescence assay showed that the proportion of Cytokeratin 7 positive cells was more than 91%, indicated the high purity of pig chorionic trophoblast cells. The results of TEM revealed that the cells had typical structural features of trophoblasts. These results indicated that our research could be used to obtain high-purity pig chorionic trophoblast cells simply and effectively by the method of trypsin-DNaseⅠdigestion and 2 density (35% and 45%, v/v) Percoll gradient centrifugation.

CLC Number:

S826.2

DONG Shu-sheng, TIAN Liang, YAN Pei-shi. Primary Culture of Villous Cytotrophoblasts Isolated from Pig Fullterm Placenta[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(1): 75-82.

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References

［1］陈昭光. PPARγ对胎盘滋养层细胞氨基酸转运体的调节及机制研究［D］. 上海：第二军医大学, 2014.
CHEN S G. PPARγ involved in the regulation of amino acid transporters in human placental syncytiotrophoblasts and its mechanism［D］. Shanghai : The Second Military Medical University, 2014. (in Chinese)
［2］程勇前, 聂青和, 周永兴, 等.人胎盘滋养层细胞的分离培养及IgG FcγRⅢ在滋养层细胞的表达［J］. 医学研究生学报, 2002, 15(2) : 105-108.
CHEN Y Q, NIE Q H, ZHOU Y X, et al. Isolation of villous cytotrophoblast from term human placenta and expression and location of IgG FcγRⅢ in human term placenta［J］. Journal of Medical Postgraduates, 2002, 15(2) : 105-108. (in Chinese)
［3］董峰.苦马豆素诱导山羊胎盘滋养层细胞凋亡的信号转导通路研究［D］. 杨凌：西北农林科技大学, 2014.
DONG F. Signal transduction pathways of apoptosis in goat placental trophoblast cells induced by swainsonine［D］. Yangling: Northwest A&F University, 2014. (in Chinese)
［4］TZSCHOPPE A,FAHLBUSCH F,SEIDEL J,et al.Dexamethasone stimulates the expression of leptin and 11βHSD2 in primary human placental trophoblastic cells［J］. Eur J Obstet Gyn R B, 2011, 156(1) : 50-55.
［5］刘慧, 刘淑英, 刘丽珍, 等. 绵羊胎盘滋养层细胞的体外培养与鉴定［J］. 中国兽医科学, 2012(3): 309-313.
LIU H, LIU S Y, LIU L Z, et al. Culture and identification of sheep trophoblast cells in vitro［J］. Veterinary Science in China, 2012(3) : 309-313. (in Chinese)
［6］KLIMAN H J, NESTLER J E, SERMASI E, et al. Purification, characterization, and in vitro differentiation of cytotrophoblasts from human term placentae［J］. Endocrinology, 1986, 118(4) : 1567-1582.
［7］王专家, 张宇飞, 刘淑英. 绵羊多核绒毛膜滋养层细胞的分离培养与鉴定［J］. 细胞与分子免疫学杂志, 2014, 30(1) : 82-86.
WANG Z J, ZHANG Y F, LIU S Y. Isolation, culture and identification of human chorionic trophoblast cells from sheep［J］. Chinese Journal of Cellular and Molecular Immunology, 2014, 30(1) : 82-86. (in Chinese)
［8］付祥龙, 张玉玲, 翟小伟, 等. 胎盘滋养层细胞分离培养研究进展［J］. 动物医学进展, 2012, 33(3) : 97-100.
FU X L, ZHANG Y L, ZHAI X W, et al. Progress of isolation and culture of placental trophoblast cells［J］. Progress in Veterinary Medicine, 2012, 33(3):97-100. (in Chinese)
［9］王海霞, 李金枝, 解其贵, 等. 滋养层细胞的分离培养与鉴定［J］. 中国临床医学, 2010, 17(5):724-726.
WANG H X, LI J Z, XIE Q G, et al. Isolation and differentiation of human first trimester trophoblasts［J］. Chinese Journal of Clinical Medicine, 2010, 17(5) : 724-726. (in Chinese)
［10］刘炀, 王旭贞, 管武太, 等. 亮氨酸对猪胎盘滋养层细胞增殖和周期的影响［J］. 中国畜牧杂志, 2016, 52(7): 35-39.
LIU Y, WANG X Z, GUAN W T, et al. Effect of leucine on proliferation and cell cycle of porcine trophectoderm cell［J］. Chinese Journal of Animal Science, 2016,52(7) : 35-39. (in Chinese)
［11］王英慧. 成纤维细胞的生物学特性及其成骨作用的研究进展［J］. 大同医学专科学校学报, 2006, 26(2):30-32.
WANG Y H. Advances in biological characteristics and osteogenesis of fibroblasts［J］. Journal of Datong Medical College, 2006, 26(2) : 30-32. (in Chinese)
［12］BLASCHITZ A, HARTMANN M, DOHR G. Marker antibodies for an efficient discrimination between trophoblast cells and other cellular components present in human first trimester placenta［J］. Placenta, 1997, 18:14.
［13］KING A, THOMAS L, BISCHOF P. Cell culture models of trophoblast ii: trophoblast cell lines— a workshop report［J］. Placenta, 2001, 21 : 104-106.
［14］HAIGH T, CHEN C P, JONES C, et al. Studies of mesenchymal cells from 1st trimester human placenta: expression of cytokeratin outside the trophoblast lineage［J］. Placenta, 1999, 20(8) : 615-625.
［15］徐浩翔, 闫言. 波形蛋白表达和功能的研究进展［J］. 北京大学学报：医学版, 2009,41(5) : 605-607.
XU H X, YAN Y. Progress of research on the expression and function of vimentin［J］. Journal of Peking University(Health Sciences), 2009, 41(5) : 605-607. (in Chinese)
［16］张小红, 李玉红, 许倩, 等. 人早孕绒毛滋养层细胞原代培养：差异贴壁法与消化排除法联合应用的可行性［J］. 中国组织工程研究与临床康复, 2010, 14(28): 5220-5223.
ZHANG X H, LI Y H, XU Q, et al. Primary culture of human chorionic trophoblast cells by differential attachment in combination with digestion elimination methods［J］. Journal of Clinical Rehabilitative Tissue Engineering Research, 2010, 14(28) : 5220-5223. (in Chinese)
［17］张久聪, 孙利, 聂青和. 人胎盘滋养层细胞原代的体外培养与改进［J］. 生物技术通讯, 2010, 21(1):58-61.
ZHANG J C, SUN L, NIE Q H. Isolation and further improvement of human trophoblasts in vitro［J］. Letters in Biotechology, 2010, 21(1): 58-61. (in Chinese)

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Primary Culture of Villous Cytotrophoblasts Isolated from Pig Fullterm Placenta

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