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Merck

Primary epithelial cell models for cystic fibrosis research.

Methods in molecular biology (Clifton, N.J.) (2011-05-07)
Scott H Randell, M Leslie Fulcher, Wanda O'Neal, John C Olsen
摘要

When primary human airway epithelial (hAE) cells are grown in vitro on porous supports at an air-liquid interface (ALI), they recapitulate in vivo morphology and key physiologic processes. These cultures are useful for studying respiratory tract biology and diseases and for testing new cystic fibrosis (CF) therapies. This chapter gives protocols enabling creation of well-differentiated primary CF and non-CF airway epithelial cell cultures with non-proprietary reagents. We also discuss the production of retroviral and lentiviral vectors, the derivation of hAE cell lines, reporter gene assays, and the evolving science of gene overexpression and knockdown in ALI hAE cultures.

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脱氧核糖核酸酶 I 来源于牛胰腺, lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein
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蛋白酶 来源于灰色链霉菌, Type XIV, ≥3.5 units/mg solid, powder
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全转铁蛋白 人, powder, BioReagent, suitable for cell culture, ≥97%
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胶原酶 来源于溶组织梭菌, suitable for release of physiologically active rat epididymal adipocytes, Type II, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
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3,3′,5-三碘-L-甲状腺氨酸 钠盐, powder, BioReagent, suitable for cell culture
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牛血清白蛋白 来源于牛血清, lyophilized powder, essentially globulin free, ≥99% (agarose gel electrophoresis)
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胶原蛋白 来源于人类胎盘, Bornstein and Traub Type IV, powder
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牛血清白蛋白 来源于牛血清, lyophilized powder, essentially IgG-free, low endotoxin, BioReagent, suitable for cell culture
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(−)-肾上腺素
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胰蛋白酶抑制剂 来源于大豆, lyophilized powder
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氢化可的松-水溶性, BioReagent, suitable for cell culture
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腺苷 5'-三磷酸 二钠盐 水合物, Grade II, ≥97% (HPLC), crystalline, from microbial
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偏硅酸钠 九水合物, BioReagent, suitable for plant cell culture, ≥98%