Spatial Control of Oxygen Delivery to Three-dimensional Cultures Alters Cancer Cell Growth and Gene Expression

Overview

Journal J Cell Physiol

Specialties Cell Biology
Physiology

Date 2019 Apr 24

PMID 31012116

Citations 9

Authors

William J Wulftange

Michelle A Rose

Marcial Garmendia-Cedillos

Davi da Silva

Joanna E Poprawski

Dhruv Srinivasachar

Taylor Sullivan

Langston Lim

Valery V Bliskovsky

Matthew D Hall

Thomas J Pohida

Robert W Robey

Nicole Y Morgan

Michael M Gottesman

Affiliations

Soon will be listed here.

Abstract

Commonly used monolayer cancer cell cultures fail to provide a physiologically relevant environment in terms of oxygen delivery. Here, we describe a three-dimensional (3D) bioreactor system where cancer cells are grown in Matrigel in modified six-well plates. Oxygen is delivered to the cultures through a polydimethylsiloxane (PDMS) membrane at the bottom of the wells, with microfabricated PDMS pillars to control oxygen delivery. The plates receive 3% oxygen from below and 0% oxygen at the top surface of the media, providing a gradient of 3-0% oxygen. We compared growth and transcriptional profiles for cancer cells grown in Matrigel in the bioreactor, 3D cultures grown in 21% oxygen, and cells grown in a standard hypoxia chamber at 3% oxygen. Additionally, we compared gene expression of conventional two-dimensional monolayer culture and 3D Matrigel culture in 21% oxygen. We conclude that controlled oxygen delivery may provide a more physiologically relevant 3D system.

Citing Articles

Preparation of Highly Functional Spheroid of Endocrine Cells Based on Thermosensitive Glycol Chitosan.

Jang S, Park Y, Huh K, Lee D Tissue Eng Regen Med. 2025; .

PMID: 39998745 DOI: 10.1007/s13770-025-00708-x.

Engineered 3D ex vivo models to recapitulate the complex stromal and immune interactions within the tumor microenvironment.

Ravi K, Manoharan T, Wang K, Pockaj B, Nikkhah M Biomaterials. 2023; 305:122428.

PMID: 38147743 PMC: 11098715. DOI: 10.1016/j.biomaterials.2023.122428.

Development and characterization of a recombinant silk network for 3D culture of immortalized and fresh tumor-derived breast cancer cells.

Collodet C, Blust K, Gkouma S, Stahl E, Chen X, Hartman J Bioeng Transl Med. 2023; 8(5):e10537.

PMID: 37693069 PMC: 10487315. DOI: 10.1002/btm2.10537.

Setup of human liver-chips integrating 3D models, microwells and a standardized microfluidic platform as proof-of-concept study to support drug evaluation.

Cox B, Barton P, Class R, Coxhead H, Delatour C, Gillent E Biomater Biosyst. 2023; 7:100054.

PMID: 36824483 PMC: 9934436. DOI: 10.1016/j.bbiosy.2022.100054.

Design considerations of benchtop fluid flow bioreactors for bio-engineered tissue equivalents .

Hoyle H, Stenger C, Przyborski S Biomater Biosyst. 2023; 8:100063.

PMID: 36824373 PMC: 9934498. DOI: 10.1016/j.bbiosy.2022.100063.

References

Mehta G, Mehta K, Sud D, Song J, Bersano-Begey T, Futai N . Quantitative measurement and control of oxygen levels in microfluidic poly(dimethylsiloxane) bioreactors during cell culture. Biomed Microdevices. 2006; 9(2):123-34. DOI: 10.1007/s10544-006-9005-7. View

Heddleston J, Li Z, McLendon R, Hjelmeland A, Rich J . The hypoxic microenvironment maintains glioblastoma stem cells and promotes reprogramming towards a cancer stem cell phenotype. Cell Cycle. 2009; 8(20):3274-84. PMC: 2825672. DOI: 10.4161/cc.8.20.9701. View

Beachley V, Wolf M, Sadtler K, Manda S, Jacobs H, Blatchley M . Tissue matrix arrays for high-throughput screening and systems analysis of cell function. Nat Methods. 2015; 12(12):1197-204. PMC: 4666781. DOI: 10.1038/nmeth.3619. View

Lewis D, Park K, Tang V, Xu Y, Pak K, Eisinger-Mathason T . Intratumoral oxygen gradients mediate sarcoma cell invasion. Proc Natl Acad Sci U S A. 2016; 113(33):9292-7. PMC: 4995943. DOI: 10.1073/pnas.1605317113. View

Bolger A, Lohse M, Usadel B . Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014; 30(15):2114-20. PMC: 4103590. DOI: 10.1093/bioinformatics/btu170. View

Riedl A, Schlederer M, Pudelko K, Stadler M, Walter S, Unterleuthner D . Comparison of cancer cells in 2D vs 3D culture reveals differences in AKT-mTOR-S6K signaling and drug responses. J Cell Sci. 2016; 130(1):203-218. DOI: 10.1242/jcs.188102. View

Chambers K, Mosaad E, Russell P, Clements J, Doran M . 3D Cultures of prostate cancer cells cultured in a novel high-throughput culture platform are more resistant to chemotherapeutics compared to cells cultured in monolayer. PLoS One. 2014; 9(11):e111029. PMC: 4224379. DOI: 10.1371/journal.pone.0111029. View

Pampaloni F, Reynaud E, Stelzer E . The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol. 2007; 8(10):839-45. DOI: 10.1038/nrm2236. View

Wulftange W, Rose M, Garmendia-Cedillos M, da Silva D, Poprawski J, Srinivasachar D . Spatial control of oxygen delivery to three-dimensional cultures alters cancer cell growth and gene expression. J Cell Physiol. 2019; 234(11):20608-20622. PMC: 6660365. DOI: 10.1002/jcp.28665. View

10.

Dobin A, Davis C, Schlesinger F, Drenkow J, Zaleski C, Jha S . STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2012; 29(1):15-21. PMC: 3530905. DOI: 10.1093/bioinformatics/bts635. View

11.

Bissell M, Radisky D, Rizki A, Weaver V, Petersen O . The organizing principle: microenvironmental influences in the normal and malignant breast. Differentiation. 2002; 70(9-10):537-46. PMC: 2933198. DOI: 10.1046/j.1432-0436.2002.700907.x. View

12.

Schneider C, Rasband W, Eliceiri K . NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012; 9(7):671-5. PMC: 5554542. DOI: 10.1038/nmeth.2089. View

13.

Samorezov J, Alsberg E . Spatial regulation of controlled bioactive factor delivery for bone tissue engineering. Adv Drug Deliv Rev. 2014; 84:45-67. PMC: 4428953. DOI: 10.1016/j.addr.2014.11.018. View

14.

El Guerrab A, Cayre A, Kwiatkowski F, Privat M, Rossignol J, Rossignol F . Quantification of hypoxia-related gene expression as a potential approach for clinical outcome prediction in breast cancer. PLoS One. 2017; 12(4):e0175960. PMC: 5400273. DOI: 10.1371/journal.pone.0175960. View

15.

Adler M, Polinkovsky M, Gutierrez E, Groisman A . Generation of oxygen gradients with arbitrary shapes in a microfluidic device. Lab Chip. 2010; 10(3):388-91. PMC: 2887752. DOI: 10.1039/b920401f. View

16.

Vaupel P, Kallinowski F, Okunieff P . Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. Cancer Res. 1989; 49(23):6449-65. View

17.

Haycock J . 3D cell culture: a review of current approaches and techniques. Methods Mol Biol. 2010; 695:1-15. DOI: 10.1007/978-1-60761-984-0_1. View

18.

Pusch J, Votteler M, Gohler S, Engl J, Hampel M, Walles H . The physiological performance of a three-dimensional model that mimics the microenvironment of the small intestine. Biomaterials. 2011; 32(30):7469-78. DOI: 10.1016/j.biomaterials.2011.06.035. View

19.

Zhang B, Radisic M . Organ-on-a-chip devices advance to market. Lab Chip. 2017; 17(14):2395-2420. DOI: 10.1039/c6lc01554a. View

20.

Grist S, Schmok J, Liu M, Chrostowski L, Cheung K . Designing a Microfluidic Device with Integrated Ratiometric Oxygen Sensors for the Long-Term Control and Monitoring of Chronic and Cyclic Hypoxia. Sensors (Basel). 2015; 15(8):20030-52. PMC: 4570408. DOI: 10.3390/s150820030. View