Worthington 肾脏组织分离指南

发布者：艾美捷科技发布时间：2022-06-07

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原代细胞分离中，针对肾脏细胞组织涉及到管段，肾小管、近端小管、肾皮质和肾细胞、肾间充质等等的解离通常会使用酶，如何选择合适的酶来完成对应肾脏细胞组织的解离，以及原代细胞采集的常规操作， 艾美捷针对肾脏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：肾脏组织分离指南

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。

肾脏
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
禽类	Avian, house sparrows	管段 / Tubule segments	Collagenase Type 2: 0.1%	HBSS	Goldstein D., Reddy V., and Plaga K.: Second Messenger Production in Avian Medullary Nephron Segments in Response to Peptide Hormones, Am J Physiol 276, R847, 1999
禽类	Avian, chicken	肾小管和肾小球 / Tubule and glomeruli	Collagenase Type 1: 0.1%	RPMI 1640	Reich, C. and Bonar, R.: Separation of Avian Kidney Tubules and Glomeruli for In Vitro Culture, Meth Cell Sci 7, 97, 1982
牛	Bovine	乳头管 / Papillary duct	Hyaluronidase: 0.2%	Keri's buffer；HEPES buffered saline	Husted, R., Hayashi, M., and Stokes, J.: Characteristics of Papillary Collecting Duct Cells in Primary Culture, Am J Physiol 255, F1160, 1988
犬/狗	Canine	肾脂肪来源的细胞 / Renal adipose derived cells	Collagenase Type 1: 0.3%	DMEM	Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011
犬/狗	Dog, 1-5 months	近端小管 / Proximal tubules	Collagenase Type 1: 0.15%	Krebs Ringer bicarbonate buffer	States, B., Reynolds, R., Lee, J., and Segal, S.: Cystine Uptake By Cultured Cells Originating From Dog Proximal Tubule Segments, In Vitro Cell Dev Biol 26, 105, 1990
犬/狗	Dog, mongrel, 20 Kg	近端管状 / Proximal tubular	Deoxyribonuclease I: 0.0125%	(see reference)	Yau, C., Rao, L., and Silverman, M.: Sugar Uptake Into a Primary Culture of Dog Kidney Proximal Tubular Cells, Can J Physiol Pharmacol 63, 417, 1985
鱼	Winter flounder (Pseudopleur-onectes americanus)	肾小管 / Renal tubule	Trypsin: 0.2%	CMF solution	Dickman, K. and Renfro, J.: Primary Culture of Flounder Renal Tubule Cells:Transepithelial Transport, Am J Physiol 251, 424, 1986
豚鼠	Guinea-pig, male, 100 g	单细胞 / Single cells	Trypsin: 0.25%	CF salt solution	Phillips, H.: Dissociation of Single Cells from Lung or Kidney Tissue with Elastase, In Vitro 8, 101, 1972
仓鼠	Hamster	乳头管 / Papillary duct	Hyaluronidase: 0.2%	Keri's buffer；HEPES buffered saline	Husted, R., Hayashi, M., and Stokes, J.: Characteristics of Papillary Collecting Duct Cells in Primary Culture, Am J Physiol 255, F1160, 1988
人	Human	肾脂肪来源的细胞 / Renal adipose derived cells	Collagenase Type 1: 0.3%	DMEM	Basu, J., Genheimer, C., Sangha, N., Quinlan, S., Guthrie, K., Kelley, R., Ilagan, R., Jain, D., Bertram, T. and Ludlow, J.: Organ Specific Regenerative Markers in Peri-Organ Adipose: Kidney., Lipids Health Dis 10, 171, 2011
人	Human, adult	肾肿瘤细胞和近端肾小管上皮 / Renal tumor cells and proximal tubular epithelial	Collagenase Type 2: 0.1%	DMEM/F12	Valente, M., Henrique, R., Costa, V., Jeronimo, C., Carvalho, F., Bastos, M., Guedes de Pinho, P. and Carvalho, M.: A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens., PLoS ONE 6, e19337, 2011
人	Human	肾细胞 / Renal cells	Collagenase Type 4: 300 u/ml； Neutral Protease: see reference	HBSS	Presnell, S., Bruce, A., Wallace, S., Choudhury, S., Genheimer, C., Cox, B., Guthrie, K., Werdin, E., Tatsumi-Ficht, P., Ilagan, R., Kelley, R., Rivera, E., Ludlow, J., Wagner, B., Jayo, M. and Bertram, T.: Isolation, Characterization, and Expansion Methods for Defined Primary Renal Cell Populations from Rodent, Canine, and Human Normal and Diseased Kidneys., Tiss Eng 17, 261, 2011
人	Human, adult	肾近曲小管和皮质成纤维细胞 / Renal proximal tubule and cortical fibroblasts	Collagenase Type 2: 383 u/ml	DMEM/F-12	Johnson, D., Saunders, H., Johnson, F., Huq, S., Field, M., and Pollock, C.: Cyclosporin exerts a direct fibrogenic effect on human tubulointerstitial cells: roles of insulin-like growth factor I, transforming growth factor beta1, and platelet-derived growth factor, J Pharmacol Exp Ther 289, 535-42, 1999
人	Human	肾皮质 / Renal cortex	Trypsin: 0.1%	Tissue Culture Grade Water	McAteer, J, Kempson, S., and Evan, A: Culture of Human Renal Cortex Epithelial Cells, J Tiss Cul Meth 13, 143, 1991
人	Human	乳头管 / Papillary duct	Collagenase: 400 u/ml	Eagle's MEM-HEPES buffer w/L-glutamine	Trifillis, A. and Kahng, M.: Characterization of an In Vitro System of Human Renal Papillary Collecting Duct Cells, In Vitro Cell Dev Biol 26, 441, 1990
人	Human	肾小球系膜 / Mesangial	Trypsin: 0.25%	DMEM/Ham's F-12	Heieren, M., van der Woude, F., and Balfour Jr., H.: Cytomegalovirus Replicates Efficiently in Human Kidney Mesangial Cells, Proc Natl Acad Sci U S A 85, 1642, 1988
人	Human, adult	管状 / Tubular	Collagenase: 250 u/ml	PBS	Yang, A., Gould-Kostka, J., and Oberley, T.: In Vitro Growth and Differentiation of Human Kidney Tubular Cells on a Basement Membrane Substrate, In Vitro Cell Dev Biol 23 (1), 34, 1987
人	Human, adult, 14-66 years	管状 / Tubular	Collagenase: 100 u/ml	Joklik's MEM	Trifillis, A., Regec, A., and Trump, B.: Isolation, Culture, and Charaterization of Human Renal Tubular Cells, J Urol 133, 324, 1985
人	Human	恶性，间质 / Malignant，Stromal	Papain: 0.009%	Sacks solution	Hemstreet, G., Enoch, P., and Pretlow, T.: Tissue Disaggregation of Human Renal Cell Carcinoma with Further Isopyknic and Isokinetic Gradient Purification, Cancer Res 40, 1043, 1980
人	Human, 24 newborn and stillborn	肾 / Renal	Trypsin: 0.25%	See reference	De Oca, H., and Malinin, T.: Dispersion and Cultivation of Renal Cells After Short-Term Storage of Kidneys, J Clin Microbiol 2, 243, 1975
猴	Monkey	肾 / Kidney	Trypsin: 0.01% - 0.00001%	Eagle's MEM	Melnick, J., and Wallis, C.: Problems Related to the Use of Serum and Trypsin in the Growth of Monkey Kidney Cells, Dev Biol 37, 77, 1976
小鼠	Mouse	巨噬细胞 / Mactrophages	Collagenase Type 4: 0.1%； Deoxyribonuclease I: 0.01%	DMEM	Yuan, Y., Li, L., Zhu, L., Liu, F., Tang, X., Liao, G., Liu, J., Cheng, J., Chen, Y. and Lu, Y.: Mesenchymal Stem Cells Elicit Macrophages into M2 Phenotype Via Improving Transcription Factor EB-Mediated Autophagy to Alleviate Diabetic Nephropathy., Stem Cells 38, 639-652, 2020
小鼠	Mouse	肾 / Kidney	Deoxyribonuclease I: 125 u/ml； Protease, Bacillus Licheniformis: 1%	DPBS	Adam, M., Potter, A. and Potter, S.: Psychrophilic Proteases Dramatically Reduce Single-Cell RNA-Seq Artifacts: A Molecular Atlas of Kidney Development., Development 144, 3625-3632, 2017
小鼠	Mouse	集合管上皮 / Collecting duct epithelial	Collagenase Type 2: 0.1%	DMEM	DiRocco, D., Kobayashi, A., Taketo, M., McMahon, A. and Humphreys, B.: Wnt4/β-catenin Signaling in Medullary Kidney Myofibroblasts., J Am Soc Nephrol 24, 1399, 2013
小鼠	Mouse	肾小球 / Glomeruli	Collagenase Type 4: 0.1%	RPMI 1640	Kabgani, N., Grigoleit, T., Schulte, K., Sechi, A., Sauer-Lehnen, S., Tag, C., Boor, P., Kuppe, C., Warsow, G., Schordan, S., Mostertz, J., Chilukoti, R., Homuth, G., Endlich, N., Tacke, F. and Weiskirchen, R.: Primary Cultures of Glomerular Parietal Epithelial Cells or Podocytes with Proven Origin., PLoS ONE 7, e34907, 2012
小鼠	Mouse	肾小球 / Glomeruli	Collagenase Type 1: 0.1%； Deoxyribonuclease I: 100 u/ml	DMEM	Sedrakyan, S., Da Sacco, S., Milanesi, A., Shiri, L., Petrosyan, A., Varimezova, R., Warburton, D., Lemley, K., De Filippo, R. and Perin, L.: Injection of Amniotic Fluid Stem Cells Delays Progression of Renal Fibrosis., J Am Soc Nephrol 23, 661-73, 2012
小鼠	Mouse, 10-14 week	近端小管 / Proximal tubule	Collagenase Type 1: 0.1%	See reference	Park, J., Pasupulati, R., Feldkamp, T., Roeser, N. and Weinberg, J.: Cyclophilin D and the Mitochondrial Permeability Transition in Kidney Proximal Tubules after Hypoxic and Ischemic Injury., Am J Physiol/Renal 301, F134, 2011
小鼠	Mouse, embryonic	肾 / Kidney	Collagenase Type 1: 0.25%； Pancreatin: 1.0%； Deoxyribonuclease I: 1 u/ml	HBSS	Brown, A., Blank, U., Adams, D., Karolak, M., Fetting, J., Hill, B. and Oxburgh, L.: Isolation and Culture of Cells from the Nephrogenic Zone of the Embryonic Mouse Kidney., J Vis Exp 50, e2555, 2011
小鼠	Mouse, adult	肾球来源 / Kidney sphere-derived	Collagenase Type 4: 0.1%	Iscove's Modified Dulbecco's	Huang, Y., Johnston, P., Zhang, B., Zakari, A., Chowdhry, T., Smith, R., Marban, E., Rabb, H. and Womer, K.: Kidney-Derived Stromal Cells Modulate Dendritic and T Cell Responses., J Am Soc Nephrol 20, 831-41, 2009