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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
要旨

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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Sigma-Aldrich
デオキシリボヌクレアーゼ I ウシ膵臓由来, lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein
Sigma-Aldrich
プロテアーゼ Streptomyces griseus由来, Type XIV, ≥3.5 units/mg solid, powder
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ホロトランスフェリン ヒト, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
コラゲナーゼ Clostridium histolyticum由来, suitable for release of physiologically active rat epididymal adipocytes, Type II, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
Sigma-Aldrich
3,3′,5-トリヨード-L-チロニン ナトリウム塩, powder, BioReagent, suitable for cell culture
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ウシ血清アルブミン ウシ血清由来, lyophilized powder, essentially IgG-free, low endotoxin, BioReagent, suitable for cell culture
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ウシ血清アルブミン ウシ血清由来, lyophilized powder, essentially globulin free, ≥99% (agarose gel electrophoresis)
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コラーゲン ヒト胎盤由来, Bornstein and Traub Type IV, powder
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(−)-エピネフリン
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ヒドロコルチゾン-水溶性, BioReagent, suitable for cell culture
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トリプシンインヒビター Glycine max(ダイズ)由来, lyophilized powder
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アデノシン 5′-三リン酸 二ナトリウム塩 水和物, Grade II, ≥97% (HPLC), crystalline, from microbial
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メタケイ酸ナトリウム 九水和物, BioReagent, suitable for plant cell culture, ≥98%