TY - JOUR

T1 - Studies on oxygen availability and the creation of natural and artificial oxygen gradients in gelatin-methacryloyl hydrogel 3D cell culture

AU - Schmitz, Carola

AU - Pepelanova, Iliyana

AU - Ude, Christian

AU - Lavrentieva, Antonina

N1 - Funding Information: This study was funded by the German Research Foundation (DFG Project 398007461 488 “3D Dual‐Gradient Systems for Functional Cell Screening” and “BiomolecularSensor Platform for Elucidating Hypoxic Signatures in 2D and 3D in vitro culture Systems”). This work was also supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany through the SMART BIOTECS alliance between the Technische Universität Braunschweig and the Leibniz Universität Hannover. Moreover, we would like to thank Dr. Janina Bahnemann and Steffen Winkler, who provided the 3D printing setup as well as Vsevolod V. Belousov and Ekaterina Potekhina for providing sensor plasmids.

PY - 2022/11

Y1 - 2022/11

N2 - Three-dimensional (3D) cultivation platforms allow the creation of cell models, which more closely resemble in vivo-like cell behavior. Therefore, 3D cell culture platforms have started to replace conventional two-dimensional (2D) cultivation techniques in many fields. Besides the advantages of 3D culture, there are also some challenges: cultivation in 3D often results in an inhomogeneous microenvironment and therefore unique cultivation conditions for each cell inside the construct. As a result, the analysis and precise control over the singular cell state is limited in 3D. In this work, we address these challenges by exploring ways to monitor oxygen concentrations in gelatin methacryloyl (GelMA) 3D hydrogel culture at the cellular level using hypoxia reporter cells and deep within the construct using a non-invasive optical oxygen sensing spot. We could show that the appearance of oxygen limitations is more prominent in softer GelMA-hydrogels, which enable better cell spreading. Beyond demonstrating novel or space-resolved techniques of visualizing oxygen availability in hydrogel constructs, we also describe a method to create a stable and controlled oxygen gradient throughout the construct using a 3D printed flow-through chamber.

AB - Three-dimensional (3D) cultivation platforms allow the creation of cell models, which more closely resemble in vivo-like cell behavior. Therefore, 3D cell culture platforms have started to replace conventional two-dimensional (2D) cultivation techniques in many fields. Besides the advantages of 3D culture, there are also some challenges: cultivation in 3D often results in an inhomogeneous microenvironment and therefore unique cultivation conditions for each cell inside the construct. As a result, the analysis and precise control over the singular cell state is limited in 3D. In this work, we address these challenges by exploring ways to monitor oxygen concentrations in gelatin methacryloyl (GelMA) 3D hydrogel culture at the cellular level using hypoxia reporter cells and deep within the construct using a non-invasive optical oxygen sensing spot. We could show that the appearance of oxygen limitations is more prominent in softer GelMA-hydrogels, which enable better cell spreading. Beyond demonstrating novel or space-resolved techniques of visualizing oxygen availability in hydrogel constructs, we also describe a method to create a stable and controlled oxygen gradient throughout the construct using a 3D printed flow-through chamber.

KW - 3D cell culture

KW - AD-MSCs

KW - hydrogels

KW - hypoxia reporter cells

KW - hypoxia sensor

KW - oxygen concentration measurements

UR - http://www.scopus.com/inward/record.url?scp=85135804241&partnerID=8YFLogxK

U2 - 10.1002/term.3344

DO - 10.1002/term.3344

M3 - Article

C2 - 35962761

AN - SCOPUS:85135804241

VL - 16

SP - 977

EP - 986

JO - Journal of Tissue Engineering and Regenerative Medicine

JF - Journal of Tissue Engineering and Regenerative Medicine

SN - 1932-6254

IS - 11

ER -