worthington 生殖相关细胞组织分离指南

发布者：艾美捷科技发布时间：2022-06-09

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原代细胞分离中，针对生殖相关细胞组织涉及到黄体细胞，黄体、成纤维细胞、上皮，子宫内膜，卵巢，子宫和羊膜间充质干细胞等等的解离通常会使用酶，如何选择合适的酶来完成对应生殖相关细胞组织的解离，以及原代细胞采集的常规操作， 艾美捷针对生殖相关细胞相关的组织分类解离分种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献，详细汇总指南：生殖相关组织细胞分离指南.png

本技术指南描述了标准实验室程序；为建立细胞分离方案提供了一种合乎逻辑的实验方法；并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南。

生殖相关细胞
种属	种属详情	细胞名称	酶组合推荐，含推荐浓度	培养基	参考文献
牛	Bovine	黄体细胞 / Corpus leuteal cells	Collagenase Type 4: 420 u/ml	M-199	Levy N, Gordin M, Mamluk R, Yanagisawa M, Smith M F, Hampton J H, Meidan R: Distinct cellular localization and regulation of endothelin-1 and endothelin-converting enzyme-1 expression in the bovine corpus luteum: implications for luteolysis, Endocrinology 142, 5254-60, 2001
牛	Bovine, female	黄体 / Leuteal	Collagenase Type 1: 0.2%	Ham's F-12	Tsang PC, Poff JP, Boulton EP, Condon WA: Four-day-old bovine corpus luteum: progesterone production and identification of matrix metalloproteinase activity in vitro, Biol Reprod 53, 1160-8, 1995
牛	Calf, mid to late gestational	成纤维细胞 / Fibroblasts	Collagenase: 0.1%	Medium 199	Coplen, D., Howard, P., Duckett, J., Snyder, H., and Macarak, E.: Characterization of a Fibroblast Cell From the Urinary Bladder Wall, In Vitro Cell Dev Biol 30A, 604, 1994
牛	Bovine	上皮，子宫内膜 / Epithelial，Endometrial	Collagenase Type 2: 0.1%	DMEM/EBSS	Munson, L., Chandler, S., and Schlafer, D.: Long-Term Culture of Bovine Trophoblastic Cells, J Tiss Cul Meth 11 (3), 123, 1988
牛	Bovine	内，黄体，子宫内膜，卵巢，子宫 / Interna & corpus luteum，Endometrium，Ovarian，Uterine	Pronase: 0.1%	Moscona's BSS	Marcus, G., Connor, L., Domingo, M., Tsang, B., Downey, B., and Ainsworth, L.: Enzymatic Dissociation of Ovarian and Uterine Tissues, Endocr Res 10, 151, 1984
犬/狗	Canine	羊膜间充质干细胞 / Amniotic membrane MSC	Trypsin: 0.25%； Collagenase Type 1: 0.2%	LG-DMEM	Park, S., Seo, M., Kim, H. and Kang, K.: Isolation and Characterization of Canine Amniotic Membrane-Derived Multipotent Stem Cells., PLoS ONE 7, e44693, 2012
鸡	Chicken, Gallus Domesticus,20-30 week	初级卵泡 / Primary follicles	Trypsin: 0.15%； Collagenase Type 1: 0.125%	Dulbecco's phosphate buffered saline	Du Meihong, Han Haitang, Jiang Bin, Zhao Chen, Qian Changsong, Shen Haiyan, Xu Yan, Li Zandong: An efficient isolation method for domestic hen (Gallus domesticus) ovarian primary follicles, J Reprod Dev 52, 569-76, 2006
鱼	Sebastes Schlegel	卵巢 / Ovarian	Collagenase Type 1: 500 u/ml	L-15	Ryu, J., Kim, H., Bae, S., Jung, C. and Gong, S.: Isolation and in Vitro Culture of Primary Cell Populations Derived from Ovarian Tissues of the Rockfish, Sebastes schlegeli, Fish Aquatic Sci 19, , 2016
青蛙	Xenopus Laevis, female	卵母细胞 / Oocytes	Collagenase Type 1: 0.2%	Barth's solution	Nicoll, G, Jawad, A., Weymouth, R, Zhang, H. and Beg, A.: Pharmacological characterization of the excitatory ‘Cys-loop’ GABA receptor family in Caenorhabditis elegans, Br J Pharmacol 174, 781, 2017
青蛙	Xenopus laevis	卵母细胞 / Oocytes	Collagenase: 0.2%	Barth's solution	Chatzidaki, A., D'Oyley, J., Gill-Thind, J., Sheppard, T. and Millar, N.: The Influence of Allosteric Modulators and Transmembrane Mutations on Desensitisation and Activation of alpha7 Nicotinic Acetylcholine Receptors., Neuropharmacol 97, 75-85, 2015
青蛙	Xenopus	卵母细胞 / Oocytes	Collagenase Type 2: 0.2%	see reference	O'Connell, D., Mruk, K., Rocheleau, J. and Kobertz, W.: Xenopus laevis Oocytes Infected with Multi-drug-resistant Bacteria: Implications for Electrical Recordings., J Gen Physiol 138, 271-7, 2011
青蛙	Xenopus laevis, female	卵母细胞 / Oocytes	Collagenase: 0.2%	see reference	Mruk, K. and Kobertz, W.: Discovery of a Novel Activator of KCNQ1-KCNE1 K Channel Complexes., PLoS ONE 4, e4236, 2009
青蛙	Xenopus laevis, female	卵母细胞 / Oocytes	Collagenase: 0.5%	Barth's solution, CF	Cohen, S., Au, S. and Pante, N.: Microinjection of Xenopus Laevis Oocytes., J Vis Exp 24, 1106, 2009
青蛙	Xenopus laevis, female	卵母细胞 / Oocytes	Collagenase Type 1: 0.2%	see reference	Pannaccione Anna, Castaldo Pasqualina, Ficker Eckhard, Annunziato Lucio, Taglialatela Maurizio: Histidines 578 and 587 in the S5-S6 linker of the human Ether-a-gogo Related Gene-1 K+ channels confer sensitivity to reactive oxygen species, J Biol Chem 277, 8912-9, 2002
青蛙	Xenopus laevis, female	卵母细胞 / Oocytes	Collagenase Type 1: 0.2%	CF Medium	Alagem, N., Dvir, M., and Reuveny, E.: Mechanism of Ba2+ Block of a Mouse Inwardly Rectifying K+ Channel: Differential Contribution by Two Discrete Residues, J Physiol 534 (2), 381, 2001
青蛙	Xenopus	卵母细胞 / Oocytes	Collagenase Type 1: 1%	(see reference)	Tian, J., Kim, S., Heilig, E., and Ruderman, J.: Identification of XPR-1, A Progesterone Receptor Required for Xenopus Oocyte Activation, Proc Natl Acad Sci U S A 97, 14358, 2000
青蛙	Xenopus laevis, female	卵母细胞 / Oocytes	Collagenase: 0.1%	Barth's solution, CF	Karkanias, N, and Papke, R.: Subtype-Specific Effects of Lithium on Glutamate Receptor Function, J Neurophysiol 81, 1506, 1999
青蛙	Xenopus laevis, mature female	卵母细胞 / Oocytes	Collagenase: 0.2%	CF Medium	Moriarty, T., Gillo, B., Carty, D., Premont, R., Landau, E., Iyengar, R.: Beta gamma Subunits of GTP-Binding Proteins Inhibit Muscarinic Receptor Stimulation of Phospholipase C, Proc Natl Acad Sci U S A 85, 8865, 1988
仓鼠	Hamster, Chinese	子房 / Ovary	Trypsin: 5%	Dialyzed fetal calf serum, 10% and 0.5M Methotrexate	Wallis, R., and Drickamer, K.: Molecular Determinants of Oligomer Formation and Complement Fixation in Mannose-Binding Proteins, J Biol Chem 274 (6), 3580, 1999
人	Human	子宫内膜上皮和间质成纤维细胞 / Endometrial epithelial and stromal fibroblasts	Collagenase Type 1: 0.32%； Hyaluronidase: 62 u/ml	DMEM	Chen, J. and Roan, N.: Isolation and Culture of Human Endometrial Epithelial Cells and Stromal Fibroblasts., Bio Protoc 5, , 2015
人	Human	子宫内膜间质 / Endometrial stromal	Collagenase Type 3: 8 u/ml； Deoxyribonuclease I: 0.01%	DMEM/F-12	Evans, J. and Salamonsen, L.: Decidualized Human Endometrial Stromal Cells are Sensors of Hormone Withdrawal in the Menstrual Inflammatory Cascade., Biol Reprod 90, 14, 2014
人	Human	内皮集落形成 / Endothelial colony forming	Collagenase Type 1: 0.1%； Deoxyribonuclease I: 0.1%； Neutral Protease: 0.075%	HBSS	Patel, J., Seppanen, E., Chong, M., Yeo, J., Teo, E., Chan, J., Fisk, N. and Khosrotehrani, K.: Prospective Surface Marker-Based Isolation and Expansion of Fetal Endothelial Colony-Forming Cells from Human Term Placenta., Stem Cells Transl Med 2, 839-47, 2013
人	Human	子宫上皮 / Uterine epithelial	Pancreatin: 0.34%； Hyaluronidase: 0.01%； Collagenase: 0.16%	HBSS	Patel, M., Ghosh, M., Fahey, J. and Wira, C.: Uterine Epithelial Cells Specifically Induce Interferon-Stimulated Genes in Response to Polyinosinic-Polycytidylic Acid Independently of Estradiol., PLoS ONE 7, e35654, 2012
人	Human	间充质干细胞 / Mesenchymal stem	Collagenase Type 1: 0.4%； Deoxyribonuclease I: 0.01%	DMEM/F12	Shalini, V., Pratheep, S., Muhammad, A., Sharmili, V., Elizabeth, G. and Rajesh, R.: Generation and Characterisation of Human Mesenchymal Stem Cells Derived From Umbilical Cord and Placenta, Regenerative Research 1, 48, 2012
人	Human	间充质干细胞 / Mesenchymal stem	Collagenase Type 2: 10%； Neutral Protease: see reference	CMF-DPBS	Steigman, S. and Fauza, D.: Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta., Curr Protoc Stem Cell Biol Chapter 1, Unit 1E.2, 2007
人	Human, female	蜕膜 / Decidual	Collagenase: 0.25%； Deoxyribonuclease I: 6.25 u/ml	DMEM/F12	Lockwood, C., Arcuri, F., Toti, P., Felice, C., Krikun, G., Guller, S., Buchwalder, L. and Schatz, F.: Tumor Necrosis Factor-Alpha and Interleukin-1 Beta Regulate Interleukin-8 Expression in Third Trimester Decidual Cells: Implications for the Genesis of Chorioamnionitis., Am J Pathol 169, 1294-302, 2006
人	Human	绒毛外滋养细胞和蜕膜基质细胞 / Extravillanous trophoblasts and decidual stromal	Trypsin: 0.1-0.25%； Collagenase Type 1: 0.1%； Deoxyribonuclease I: 0.02-0.05%	HBSS RPMI	Spessotto, P., Bulla, R., Danussi, C., Radillo, O., Cervi, M., Monami, G., Bossi, F., Tedesco, F., Doliana, R. and Colombatti, A.: EMILIN1 Represents a Major Stromal Element Determining Human Trophoblast Invasion of the Uterine Wall., J Cell Sci 119, 4574, 2006
人	Human	支持细胞 / Sertoli cells	Trypsin: 2.5%； Collagenase Type 1: 2%； Hyaluronidase: 1%	DMEM/F-12	Teng Yan, Xue Wu-jun, Ding Xiao-ming, Feng Xin-shun, Xiang He-li, Jiang Ya-zhuo, Tian Pu-xun: Isolation and culture of adult Sertoli cells and their effects on the function of co-cultured allogeneic islets in vitro, Chin Med J (Engl) 118, 1857-62, 2005
人	Human, female	子宫上皮 / Uterine epithelial	Pancreatin: 0.34%； Hyaluronidase: 0.01%； Collagenase: 0.16%	HBSS	Meter, R., Wira, C. and Fahey, J.: Secretion of Monocyte Chemotactic Protein-1 by Human Uterine Epithelium Directs Monocyte Migration in Culture., Fertil Steril 84, 191, 2005
人	Human, female, 34-51 yr	子宫内膜上皮和基质细胞 / Endometrial epithelial and stromal cells	Collagenase Type 3: 0.03%； Deoxyribonuclease I: 0.004%	DMEM/F-12	Chan Rachel W S, Schwab Kjiana E, Gargett Caroline E: Clonogenicity of human endometrial epithelial and stromal cells, Biol Reprod 70, 1738-50, 2004