Matrigel® as 3D culture medium: 2D vs. 3D changes in proliferation and ultrastructure

Alexandra A Iankoulska¹ and Richard W Cole*^2,3

DOI: 10.26717/BJSTR.2017.01.000140

Received: June 08, 2017   Published: June 16, 2017

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Abstract

3D cell culture has great potential in the field of tissue engineering and regenerative medicine. Three- dimensional ECM-like substances such as Matrigel® mimic natural proteins found in the ECM and result in a more accurate reflection of cell growth within the human body. In our lab, we compared HT1080 cells transfected with the histone H2B-green fluorescent protein (GFP) growing directly on glass (2D) to those in Matrigel® on the basis of cell growth, proliferation, division and morphology. During cell counting, the cells were maintained at a near-homeostatic temperature, humidity and CO2 level in a custom stage top environmental chamber [1] while being imaged on an Olympus (Melville, NY) IX70 inverted fluorescence microscope. Our results indicated that cell growth, division and proliferation were of greater magnitude in Matrigel® than on a glass coverslip (control). For ultrastructure imaging purposes, a culture of MDCK cells (Canis familiaris, kidney, normal) that were transfected with a pEYFP-Tubulin plasmid subcellular localization vector were used.

Widefield epifluorescent analysis indicated that there was no significant difference in cellular ultrastructure on the basis of flat/rounded morphology and microtubule distribution between the cells grown in glass-bottomed Petri dishes and those in Matrigel®. These results are consistent with our hypothesis that cells in Matrigel® would exhibit greater cell growth, proliferation, and division while maintaining normal cell morphology.

Keywords: Imaging; Fluorescence; Tissue-culture

Introduction | Materials and Methods | Results | Discussion | Acknowledgement | References | Figures | Tables |