This protocol describes the thawing, culturing, and cryopreservation of the human alveolar-like epithelial cell line NCI-H441. NCI-H441 cells should be used in place of the adenocarcinoma cell line, A549, when studying the human alveolar epithelium as, in contrast to A549 cells, H441 cells reflect the in vivo alveolar epithelium barrier and transporter protein function (Ren et al., 2016; Hermanns et al., 2004). Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.NOTE: There is a consolidated PDF version of this protocol in the supplementary files section below.
This protocol describes the thawing, culturing, and cryopreservation of the human lung microvascular endothelial cell line, HULEC-5a (Mehta et al., 2006). Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use. NOTE: There is a consolidated PDF version of this protocol in the supplementary files section below.
This protocol describes the thawing, culturing, and cryopreservation of the human lung fibroblast cell line, IMR90, in Advanced RPMI. Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use. NOTE: There is a consolidated PDF version of this protocol in the supplementary files section below.
This protocol describes the use of the ZYMO Quick-RNA miniprep kit for the isolation and purification of total RNA from cells grown in culture. The conditions described below were developed for the isolation of RNA from H441 and HULEC from the alveolar-capillary region exposure (ACRE) model. Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use. NOTE: There is a consolidated PDF version of this protocol in the supplementary files section below.
This exposure model was designed as an in vitro representation of trans-alveolar effects of airway particulate matter exposures on microvascular endothelial cells in the alveolar capillary region of the lung. The alveolar epithelial cells are represented by the NCI-H441 cell line, interstitial fibroblasts are represented by the IMR90 cell line, and microvascular endothelial cells are represented by the HULEC cell line. Disclaimer: The contents of this article have been reviewed by the US Environmental Protection Agency and approved for publication and do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use. NOTE: There is a consolidated PDF version of this protocol in the supplementary files section below.
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