Medium throughput breathing human primary cell alveolus-on-chip model

Abstract

Organs-on-chips have the potential to improve drug development efficiency and decrease the need for animal testing. For the successful integration of these devices in research and industry, they must reproduce in vivo contexts as closely as possible and be easy to use. Here, we describe a ‘breathing’ lung-on-chip array equipped with a passive medium exchange mechanism that provide an in vivo-like environment to primary human lung alveolar cells (hAEpCs) and primary lung endothelial cells. This configuration allows the preservation of the phenotype and the function of hAEpCs for several days, the conservation of the epithelial barrier functionality, while enabling simple sampling of the supernatant from the basal chamber. In addition, the chip design increases experimental throughput and enables trans-epithelial electrical resistance measurements using standard equipment. Biological validation revealed that human primary alveolar type I (ATI) and type II-like (ATII) epithelial cells could be successfully cultured on the chip over multiple days. Moreover, the effect of the physiological cyclic strain showed that the epithelial barrier permeability was significantly affected. Long-term co-culture of primary human lung epithelial and endothelial cells demonstrated the potential of the lung-on-chip array for reproducible cell culture under physiological conditions. Thus, this breathing lung-on-chip array, in combination with patients’ primary ATI, ATII, and lung endothelial cells, has the potential to become a valuable tool for lung research, drug discovery and precision medicine.

Second-generation lung-on-a-chip array with a stretchable biological membrane

“…The CE-membrane has great versatility as thickness can easily be tuned by adapting the volume of the CE solution pipetted onto the gold mesh to suit any number of experimental requirements. The thinnest membrane obtained has a thickness comparable to the thinnest porous PDMS membrane reported thus far 10 . Unlike synthetic polymers, such as PDMS, the optically transparent CE-membrane does not require any preliminary coating prior to cell seeding.…”

“…Primary hAEpCs were isolated from patient tissue according to a protocol reported previously 10,40 . Briefly, alveolar epithelial type II (AT II) cells were isolated from tissue obtained from healthy areas removed from patients undergoing lung tumor resection surgery.…”

“…Micro-engineered systems with an integrated membrane in a microfluidic setting have been reported to model various barrier tissue interfaces, such as those of the lung alveoli, the brain and the gut 7 . By implementing a flexible polymeric membrane in such microfluidic systems, mechanical forces, such as those induced by breathing, could be reproduced 8,9,10 .…”

Toxicological evaluation of airborne particulate matter. Are cell culture technologies ready to replace animal testing?

“…Research and development in the area of lung‐on‐chip technology has recently been applied to drug screening (Benam et al, ; Skardal et al, ) and to the study of lung physiology (Benam et al, ; Henry et al, ; Humayun, Chow, & Young, ; J. D. Stucki et al, ) and pathophysiology (Huh et al, ; Jain et al, ), and we wonder whether this technology was also used in investigations related to environmental toxicology. However, although the technology is promising and does not have ethical and regulatory limitations, its use to study the response of human cells to ambient toxicants has not yet been presented in the literature (Cho & Yoon, ).…”

“…Research and development in the area of lung-on-chip technology has recently been applied to drug screening (Benam et al, 2016;Skardal et al, 2017) and to the study of lung physiology (Benam et al, 2017;Henry et al, 2017;Humayun, Chow, & Young, 2018;J. D. Stucki et al, 2018) and pathophysiology (Huh et al, 2012;Jain et al, 2018), and we wonder whether this technology was also used in investigations related to environmental toxicology.…”