TY - JOUR

T1 - Towards a microfluidic H295R steroidogenesis assay—biocompatibility study and steroid detection on a thiol-ene-based chip

AU - Despicht, Caroline

AU - Munkboel, Cecilie H.

AU - Chou, Hua Nee

AU - Ertl, Peter

AU - Rothbauer, Mario

AU - Kutter, Jörg P.

AU - Styrishave, Bjarne

AU - Kretschmann, Andreas

N1 - Funding Information: We extend our sincere appreciation to Drago Sticker for sharing his expertise in the field of lab-on-a-chip technology and providing invaluable input during the initial stages of this project. We would like to thank the Independent Research Fund Denmark (project ID: DFF—7017-00267) and the Carlsberg Foundation (project no: CF18-0995) for funding this project. Publisher Copyright: © 2023, The Author(s).

PY - 2023

Y1 - 2023

N2 - The development of cell-based microfluidic assays offers exciting new opportunities in toxicity testing, allowing for integration of new functionalities, automation, and high throughput in comparison to traditional well-plate assays. As endocrine disruption caused by environmental chemicals and pharmaceuticals represents a growing global health burden, the purpose of the current study was to contribute towards the miniaturization of the H295R steroidogenesis assay, from the well-plate to the microfluidic format. Microfluidic chip fabrication with the established well-plate material polystyrene (PS) is expensive and complicated; PDMS and thiol-ene were therefore tested as potential chip materials for microfluidic H295R cell culture, and evaluated in terms of cell attachment, cell viability, and steroid synthesis in the absence and presence of collagen surface modification. Additionally, spike-recovery experiments were performed, to investigate potential steroid adsorption to chip materials. Cell aggregation with poor steroid recoveries was observed for PDMS, while cells formed monolayer cultures on the thiol-ene chip material, with cell viability and steroid synthesis comparable to cells grown on a PS surface. As thiol-ene overall displayed more favorable properties for H295R cell culture, a microfluidic chip design and corresponding cell seeding procedure were successfully developed, achieving repeatable and uniform cell distribution in microfluidic channels. Finally, H295R perfusion culture on thiol-ene chips was investigated at different flow rates (20, 10, and 2.5 µL/min), and 13 steroids were detected in eluting cell medium over 48 h at the lowest flow rate. The presented work and results pave the way for a time-resolved microfluidic H295R steroidogenesis assay. Graphical abstract: [Figure not available: see fulltext.].

AB - The development of cell-based microfluidic assays offers exciting new opportunities in toxicity testing, allowing for integration of new functionalities, automation, and high throughput in comparison to traditional well-plate assays. As endocrine disruption caused by environmental chemicals and pharmaceuticals represents a growing global health burden, the purpose of the current study was to contribute towards the miniaturization of the H295R steroidogenesis assay, from the well-plate to the microfluidic format. Microfluidic chip fabrication with the established well-plate material polystyrene (PS) is expensive and complicated; PDMS and thiol-ene were therefore tested as potential chip materials for microfluidic H295R cell culture, and evaluated in terms of cell attachment, cell viability, and steroid synthesis in the absence and presence of collagen surface modification. Additionally, spike-recovery experiments were performed, to investigate potential steroid adsorption to chip materials. Cell aggregation with poor steroid recoveries was observed for PDMS, while cells formed monolayer cultures on the thiol-ene chip material, with cell viability and steroid synthesis comparable to cells grown on a PS surface. As thiol-ene overall displayed more favorable properties for H295R cell culture, a microfluidic chip design and corresponding cell seeding procedure were successfully developed, achieving repeatable and uniform cell distribution in microfluidic channels. Finally, H295R perfusion culture on thiol-ene chips was investigated at different flow rates (20, 10, and 2.5 µL/min), and 13 steroids were detected in eluting cell medium over 48 h at the lowest flow rate. The presented work and results pave the way for a time-resolved microfluidic H295R steroidogenesis assay. Graphical abstract: [Figure not available: see fulltext.].

KW - Biocompatibility

KW - Cell-based assay

KW - Microfluidics

KW - PDMS

KW - Steroidogenesis

KW - Thiol-ene

U2 - 10.1007/s00216-023-04816-2

DO - 10.1007/s00216-023-04816-2

M3 - Journal article

C2 - 37438566

AN - SCOPUS:85164493625

VL - 415

SP - 5421

EP - 5436

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

SN - 1618-2642

ER -