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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 segmentsCollagenase Type 2: 0.1%HBSSGoldstein 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 glomeruliCollagenase Type 1: 0.1%RPMI 1640Reich, C. and Bonar, R.: Separation of Avian Kidney Tubules and Glomeruli for In Vitro Culture, Meth Cell Sci 7, 97, 1982
Bovine乳头管 / Papillary ductHyaluronidase: 0.2%Keri's buffer;HEPES buffered salineHusted, 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 cellsCollagenase Type 1: 0.3%DMEMBasu, 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 tubulesCollagenase Type 1: 0.15%Krebs Ringer bicarbonate bufferStates, 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 tubularDeoxyribonuclease 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
Winter flounder (Pseudopleur-onectes americanus)肾小管 / Renal tubuleTrypsin: 0.2%CMF solutionDickman, 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 cellsTrypsin: 0.25%CF salt solutionPhillips, H.: Dissociation of Single Cells from Lung or Kidney Tissue with Elastase, In Vitro 8, 101, 1972
仓鼠Hamster乳头管 / Papillary ductHyaluronidase: 0.2%Keri's buffer;HEPES buffered salineHusted, R., Hayashi, M., and Stokes, J.: Characteristics of Papillary Collecting Duct Cells in Primary Culture, Am J Physiol 255, F1160, 1988
Human肾脂肪来源的细胞 / Renal adipose derived cellsCollagenase Type 1: 0.3%DMEMBasu, 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
Human, adult肾肿瘤细胞和近端肾小管上皮 / Renal tumor cells and proximal tubular epithelialCollagenase Type 2: 0.1%DMEM/F12Valente, 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
Human肾细胞 / Renal cellsCollagenase Type 4: 300 u/ml;
Neutral Protease: see reference
HBSSPresnell, 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
Human, adult肾近曲小管和皮质成纤维细胞 / Renal proximal tubule and cortical fibroblastsCollagenase Type 2: 383 u/mlDMEM/F-12Johnson, 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
Human肾皮质 / Renal cortexTrypsin: 0.1%Tissue Culture Grade WaterMcAteer, J, Kempson, S., and Evan, A: Culture of Human Renal Cortex Epithelial Cells, J Tiss Cul Meth 13, 143, 1991
Human乳头管 / Papillary ductCollagenase: 400 u/mlEagle's MEM-HEPES buffer w/L-glutamineTrifillis, 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
Human肾小球系膜 / MesangialTrypsin: 0.25%DMEM/Ham's F-12Heieren, 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
Human, adult管状 / TubularCollagenase: 250 u/mlPBSYang, 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
Human, adult, 14-66 years管状 / TubularCollagenase: 100 u/mlJoklik's MEMTrifillis, A., Regec, A., and Trump, B.: Isolation, Culture, and Charaterization of Human Renal Tubular Cells, J Urol 133, 324, 1985
Human恶性,间质 / Malignant,StromalPapain: 0.009%Sacks solutionHemstreet, 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
Human, 24 newborn and stillborn肾 / RenalTrypsin: 0.25%See referenceDe Oca, H., and Malinin, T.: Dispersion and Cultivation of Renal Cells After Short-Term Storage of Kidneys, J Clin Microbiol 2, 243, 1975
Monkey肾 / KidneyTrypsin: 0.01% - 0.00001%Eagle's MEMMelnick, 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 / MactrophagesCollagenase Type 4: 0.1%;
Deoxyribonuclease I: 0.01%
DMEMYuan, 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 / KidneyDeoxyribonuclease I: 125 u/ml;
Protease, Bacillus Licheniformis: 1%
DPBSAdam, 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 epithelialCollagenase Type 2: 0.1%DMEMDiRocco, 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 / GlomeruliCollagenase Type 4: 0.1%RPMI 1640Kabgani, 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 / GlomeruliCollagenase Type 1: 0.1%;
Deoxyribonuclease I: 100 u/ml
DMEMSedrakyan, 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 tubuleCollagenase Type 1: 0.1%See referencePark, 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 / KidneyCollagenase Type 1: 0.25%;
Pancreatin: 1.0%;
Deoxyribonuclease I: 1 u/ml
HBSSBrown, 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-derivedCollagenase Type 4: 0.1%Iscove's Modified Dulbecco'sHuang, 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


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