Culture of HULEC-5a Cells

Abstract

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.

Characterizing the extracellular vesicle proteomic landscape of the human airway using in vitro organotypic multi-cellular models

“…Detailed cell culture methods for the H441 cell line is available through the online, publicly available Nature Portfolio, Protocol Exchange. 69 , 70 , 71 , 72 In brief, IMR90 were maintained in Advanced-RPMI growth medium (A-RPMI; ThermoFisher #12633020) supplemented with 5% FBS (ThermoFisher, #16000044, certified), 0.5% Penicillin/Streptomycin (P/S; ThermoFisher, #15140122) and 4 mM GlutaMAX (ThermoFisher #35050061)). Fibroblasts: IMR90 cells (human, female, lung fibroblasts; ATCC #CCL-182, Batch #64155514) represented the fibroblasts and were obtained from ATCC.…”

“…Detailed cell culture methods for the IMR90 cell line is available through the online, publicly available Nature Portfolio, Protocol Exchange. 69 , 70 , 71 , 72 In brief, IMR90 were maintained in Advanced-RPMI growth medium (A-RPMI; ThermoFisher #12633020) supplemented with 5% FBS (ThermoFisher, #16000044, certified), 0.5% Penicillin/Streptomycin (P/S; ThermoFisher, #15140122) and 4 mM GlutaMAX (ThermoFisher #35050061)). Lung microvascular endothelial cells: HULEC-5a cells (hereafter referred to as “HULEC”; human, male, lung microvascular endothelial cells, ATCC #CRL-3244, Batch #70025430) represented the endothelial cells and were obtained from ATCC.…”

“…Detailed cell culture methods for the HULEC-5a cell line is available through the online, publicly available Nature Portfolio, Protocol Exchange. 69 , 70 , 71 , 72 HULEC were maintained in complete HULEC growth medium (MCDB-131 (Gibco #10372019) supplemented with 10% FBS, 1% P/S, 10 mM GlutaMAX, 10 ng/mL human Epidermal Growth Factor (Fisher Scientific, #PHG0311), and 1μg/mL hydrocortisone (MilliporeSigma, #H0888)).…”