Isolation and Purification of Mouse Spermatogonia

doi:10.3969/j.issn.1674-5817.2015.01.009

Abstract

Abstract: Objective To establish a simple and efficient method for separation and purification of mouse spermatogonia. Methods Spermatogonia were isolated and purified from neonatal mouse testis using collagenase IV/DNase I/dispase combination enzymatic digestion and differential velocity of cell adherence method. Isolated spermatogonia were analyzed by morphology and stra8 immunochemistry. Results The isolated spermatogonia were stra8 positive and cell purity was as high as 86%. Conclusion This method is efficient for separation and purification of mouse spermatogonia .

Key words: Mouse, Spermatogonia, Isolation, Purification

CLC Number:

R321.1Q95-33

LI Yu-hua, DUAN Fei, ZHOU Xiao-yu, Li Yang, LI Run-sheng. Isolation and Purification of Mouse Spermatogonia[J]. Laboratory Animal and Comparative Medicine, 2015, 35(1): 42-45.

References

[1] Bucci LR, Brock WA, Johnson TS, et al.Isolation and biochemical studies of enriched populations of spermatogonia and early primary spermatocytes from rat testes[J]. Biol Reprod, 1986, 34(1):195-206.
[2] 贺亚南, 陈晓丽, 任晓霞, 等. 免疫磁珠纯化小鼠精原干细胞的研究[J]. 中国生物工程杂志, 2014, 34(7):38-43.
[3] Lassalle B, Ziyyat A, Testart J, et al.Flow cytometric method to isolate round spermatids from mouse testis[J]. Hum Reprod, 1999, 14(2):388-394.
[4] Guan K, Wolf F, Becker A, et al.Isolation and cultivation of stem cells from adult mouse testes[J]. Nat Protoc, 2009, 4(2):143-154.
[5] 刘丹, 夏娟, 刘欢欢, 等. 哺乳动物精原干细胞体外分离培养研究进展[J]. 四川动物, 2014, 33(2):315-320.
[6] Bellve AR, Cavicchia JC, Millette CF, et al.Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization[J]. J Cell Biol, 1977, 74(1):68-85.
[7] Boucheron C and Baxendale V, Isolation and purification of murine male germ cells[J]. Methods Mol Biol, 2012. 825:59-66.
[8] Craviso GL, Generation of functionally competent single bovine adrenal chromaffin cells from cell aggregates using the neutral protease dispase[J]. J Neurosci Methods, 2004. 137(2) : 275-281.
[9] 师如意, 卫恒习, 李莉, 等. 猪精原干细胞体外培养条件的优化[J]. 华南农业大学学报, 2013,34(2):224-229.
[10] Oulad-Abdelghani M, Bouillet P, Decimo D, et al.Characterization of a premeiotic germ cell-specific cytoplasmic protein encoded by Stra8, a novel retinoic acid-responsive gene[J]. J Cell Biol, 1996, 135(2):469-477.
[11] 张晓丽, 高英茂, 赵舒武, 等. 新生小鼠精原细胞分离和纯化的实验研究[J]. 山东大学学报(医学版), 2005, 43(8): 674-677.
[12] 王永彬, 刘凤美, 张军令, 等. 小鼠精原干细胞的分离纯化及初步鉴定[J]. 黑龙江动物繁殖, 2014, 22(4):19-22.

[1]	Yongqiang NIE, Zhaoxia WANG. Rescue Technology and Its Application of Endangered Gene-Edited Mice [J]. Laboratory Animal and Comparative Medicine, 2023, 43(6): 636-640.
[2]	Ying HUANG, Siyu WEI, Li CAI, Sujing QIANG, Dongting LI, Yuqiang DING. Microbiological Monitoring Analysis of Laboratory Rats and Mice from Vendors: Department of Laboratory Animal Science of Fudan University as an Example [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 347-354.
[3]	Xue WANG, Yonghe HU. Analysis of Common Types and Construction Elements of Diabetic Mouse Models [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 415-421.
[4]	Hui HUANG, Yasheng DENG, Tianwei LIANG, Yiqing ZHENG, Yanping FAN, Na RONG, Jiang LIN. Evaluation and Analysis of Modeling Methods for Animal Models with Diminished Ovarian Reserve [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 422-428.
[5]	Fangni LIU, Junping LU, Yuehua KE, Changjun WANG, Jinpeng GUO. Prevalence of Mouse Norovirus in Experimental Mice in Beijing [J]. Laboratory Animal and Comparative Medicine, 2023, 43(2): 205-212.
[6]	Chenyin REN, Siqi GAO, Hao LI, Biao TANG, Hua YANG, Yuehuan LIU. In Vivo Colonization Test of Two CRISPR-Engineered Escherichiacoli in Mice [J]. Laboratory Animal and Comparative Medicine, 2022, 42(6): 541-550.
[7]	Feng WEI, Weiwei CHENG, Yafu YIN. Characteristics and Application of Transgenic Mouse Models in Alzheimer's Disease [J]. Laboratory Animal and Comparative Medicine, 2022, 42(5): 432-439.
[8]	Shiyan YU. Advances in the Application of Mouse Models to Study Digestive Mucosal Immunity and Infectious Diseases [J]. Laboratory Animal and Comparative Medicine, 2022, 42(1): 3-10.
[9]	LAI Suomei, DING Yifu, LI Jinsong. A New Strategy for Constructing Mouse Models of Complex Diseases: Semi-cloning Technology Based on Sperm-like Haploid Embryonic Stem Cells [J]. Laboratory Animal and Comparative Medicine, 2021, 41(5): 369-383.
[10]	SHI Meiyan, WANG Xuan, WANG Wenguang, RUAN Leiying, DAI Jiejie. Isolation and Culture of Spinal Microvascular Endothelial Cells of Tree Shrews and Experimental Study on Infection with Enterovirus 71 [J]. Laboratory Animal and Comparative Medicine, 2021, 41(2): 148-154.
[11]	YAO Ding, ZHOU Jing, YAN Guofeng, WANG Huiyang, WANG Yadi, MA Zhengwen. Establishment of Salt-sensitive Hypertension Model in C57BL/6J Mice [J]. Laboratory Animal and Comparative Medicine, 2020, 40(4): 314-.
[12]	CHAI Wenjun, SUN Lei, LIU Xiaoli, PAN Hongyu, GUO Tianan, XU Ye, YAN Mingxia. Establishment of Bone Metastasis Mouse Models through Injecting Human Lung Cancer Cells into Left Ventricle#br# under Ultrasound Guidance#br# [J]. Laboratory Animal and Comparative Medicine, 2020, 40(3): 183-.
[13]	WANG Ji, WANG Shasha, FU Rui, WANG Shujing, LI Wei, QIN Xiao, HUANG Zongwen, LI Xiaobo, GONG Wei, YUE Bingfei, HE Zhengming. Establishment of Real-time Fluorescent Quantitative PCR Method for Mouse Hepatitis Virus and Application in Some Rodents [J]. Laboratory Animal and Comparative Medicine, 2020, 40(2): 95-103.
[14]	JIANG Hongli, MA Hongye, XUE Jinhong, SUN Lingshuang, CHEN Lei. The Role of Villin-1 in Model of Habu Nephritis Mice with Unilateral Nephrectomy [J]. Laboratory Animal and Comparative Medicine, 2020, 40(1): 1-8.
[15]	ZOU Lu, DAI Tian-e, LU Xiao, AI Xiao-jie. Anesthetic Effect Evaluation of Combined Zoletil 50 with Dexdomitor in BALB/c Mouse [J]. Laboratory Animal and Comparative Medicine, 2019, 39(4): 310-313.