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使用逆转录病毒载体将基因导入大鼠气道上皮细胞。

Gene transfer into rat airway epithelial cells using retroviral vectors.

作者信息

Stanley C, Rosenberg M B, Friedmann T

机构信息

Department of Pediatrics, University of California School of Medicine, La Jolla 92093-0634.

出版信息

Somat Cell Mol Genet. 1991 Mar;17(2):185-90. doi: 10.1007/BF01232975.

Abstract

Primary cultures of epithelial cells from adult rat tracheas were maintained in vitro on collagen matrices and were exposed to a murine retrovirus vector expressing the E. coli beta-galactosidase gene. Infection was carried out on cells grown as monolayers under medium and on cells grown on raised platforms. Cells maintained at an air-medium interface were highly susceptible to infection with the vector, showing an efficiency of infection of 20-25%, compared with an efficiency of less than 1% for cells grown under medium. Infected beta-galactosidase-expressing cells were seeded into denuded tracheas and were capable of partially repopulating the denuded tracheas grafted subcutaneously into host rats. The susceptibility of these cells to retroviral infection suggests an approach to the treatment of some pulmonary genetic disorders such as cystic fibrosis.

摘要

成年大鼠气管上皮细胞的原代培养物在体外胶原基质上维持,并暴露于表达大肠杆菌β-半乳糖苷酶基因的鼠逆转录病毒载体。在培养基下单层生长的细胞以及在升高平台上生长的细胞上进行感染。维持在气-液界面的细胞对载体感染高度敏感,感染效率为20%-25%,而在培养基下生长的细胞感染效率低于1%。将感染后表达β-半乳糖苷酶的细胞接种到去上皮的气管中,它们能够部分重新填充皮下移植到宿主大鼠体内的去上皮气管。这些细胞对逆转录病毒感染的敏感性提示了一种治疗某些肺部遗传性疾病如囊性纤维化的方法。

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