worthington 垂体组织分离指南

原代细胞分离中,针对垂体组织涉及到垂体,滤泡,生长激素、垂体前叶和结节等等的解离通常会使用酶,如何选择合适的酶来完成对应垂体组织的解离,以及原代细胞采集的常规操作, 艾美捷针对垂体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:

worthington 胰腺组织分离指南

 

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

 

垂体
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
Calf, male, less then 6 months old 垂体 / Pituitary Collagenase: 0.3% DMEM Hassan, H., and Merkel, R.: Perifusion Model System to Culture Bovine Hypothalamic Slices In Series with Dispersed Anterior Pituitary Cells, In Vitro Cell Dev Biol 30A, 435, 1994
Bovine 垂体 / Pituitary Collagenase: 0.1% EBSS, CMF Mason, W. and Ingram, C.: Techniques for Studying the Role of Electrical Activity in Control of Secretion by Normal Anterior Pituitary Cells, Vol. 124,, , 207, 1986
Bovine 滤泡,垂体前叶和结节 / Follicular, anterior pituitary and pars tuberalis Deoxyribonuclease I: 200 /ml HBSS, CMF Ferrara, N., Goldsmith, P., Fujii, D., and Weiner, R.: Culture and Characterization of Follicular Cells of the Bovine Anterior Pituitary and Pars Tuberalis, Vol. 124,, , 245, 1986
Calf, male, 1-6 week old 垂体 / Pitituary Hyaluronidase: 0.1% DMEM Ridgway, E., Klibanski, A., Marorana, M., Milbury, P., Kieffer, J., and Chin, W.: The Effect of Somatostatin on the Release of Thyrotropin and its Subunits from Bovine Anterior Pitituary Cells in Vitro, Endocrinology 112 (6), 1937, 1983
小鼠 Mouse 垂体 / Pituitary Collagenase Type 2: 0.5%;
Deoxyribonuclease I: 0.005%
HBSS Perez Millain, M., Brinkmeier, M., Mortensen, A. and Camper, S.: PROP1 Triggers Epithelial-Mesenchymal Transition-Like Process in Pituitary Stem Cells., Elife 5, , 2016
小鼠 Mouse 垂体 / Pituitary Collagenase Type 1: 1,000 u/ml;
Soybean Trypsin Inhibitor: 0.01%;
Hyaluronidase: 0.1%;
Deoxyribonuclease I: 0.001%
DMEM Pyczek, J., Buslei, R., Schult, D., Holsken, A., Buchfelder, M., Hess, I., Hahn, H. and Uhmann, A.: Hedgehog Signaling Activation Induces Stem Cell Proliferation and Hormone Release in the Adult Pituitary Gland., Sci Rep 6, 24928, 2016
小鼠 Mouse, male 垂体 / Pituitary Collagenase: 0.4%;
Hyaluronidase: 0.1%;
Trypsin: 0.3%
DMEM/Han's F12 Steveson Tami C, Ciccotosto Giuseppe D, Ma Xin-Ming, Mueller Gregory P, Mains Richard E, Eipper Betty A: Menkes protein contributes to the function of peptidylglycine alpha-amidating monooxygenase, Endocrinology 144, 188-200, 2003
绵羊 Ovine, adult 生长激素 / Somatotropes Collagenase Type 1: 0.3%;
Hyaluronidase:
Medium 199 Xu Ruwei, Wang Qinling, Yan Ming, Hernandez Maria, Gong Changhong, Boon WahChin, Murata Yoko, Ueta Yoichi, Chen Chen: Orexin-A augments voltage-gated Ca2+ currents and synergistically increases growth hormone (GH) secretion with GH-releasing hormone in primary cultured ovine somatotropes, Endocrinology 143, 4609-19, 2002
大鼠 Rat, male, 12-15 week 垂体 / Pituitary Collagenase Type 2: 0.4%;
Deoxyribonuclease I: 0.04%
DMEM Akieda-Asai, S., Zaima, N., Ikegami, K., Kahyo, T, Yao, I., Hatanaka, T., Iemura, S., Sugiyama, R., Yokozeki, T., Eishi, Y., Koike, M., Ikeda, K., Chiba, T., Yamaza, H., Shimokawa, I., Song, S., Matsuno, A., Mizutani, A., Sawabe, M. Chao, N., Tanaka, M., : SIRT1 Regulates Thyroid-Stimulating Hormone Release by Enhancing PIP5K gamma Activity through Deacetylation of Specific Lysine Residues in Mammals., PLoS ONE 5, e11755, 2010
大鼠 Rat, Wistar, male, 2 month old 垂体 / Pituitary Trypsin: 0.1% DMEM Zhou, X., De Schepper, J., De Craemer, D., Delhase, M., Gys, G., Smitz, J., and Hooghe-Peters, E.: Pituitary Growth Hormone Release and Gene Expression in Cafeteria-Diet-Induced Obese Rats, J Endocrinol 159, 165, 1998
大鼠 Rat, SD, female, 200 - 250 g 垂体前叶 / Anterior pituitary gland Trypsin: 0.1% EBSS, CMF D'Emden, M. and Wark, J.: Culture Requirements for Optimal Expression of 1,25-Dihydroxyvitamin D3-Enhanced Thyrotropin Secretion, In Vitro Cell Dev Biol 27, 197, 1991
大鼠 Rat 垂体 / Pituitary Trypsin: 0.3 % HEPES Wilfinger, W., Larsen, W., Downs, T., and Wilbur, D.: An In Vitro Model for Studies of Intercellular Communication in Cultured Rat Anterior Pituitary Cells, Tissue Cell 16 (4), 483, 1984
大鼠 Rat, SD, male, 250-450 g 垂体前叶 / Anterior pituitary Trypsin: 0.25% Krebs Portanova, R., Smith, D., and Sayers, G.: A Trypsin for the Preparation of Isolated Rat Anterior Pituitary, Proc Soc Exp Biol Med 133, 573, 1970

 

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