Fabrication of co-culture device for biological applications using 3D printing: Comparison of Fused Deposition Modeling (FDM) and Stereolithography (SLA)

Harsha, Kurre Sri and Giri, Lopamudra (2019) Fabrication of co-culture device for biological applications using 3D printing: Comparison of Fused Deposition Modeling (FDM) and Stereolithography (SLA). Masters thesis, Indian institute of technology Hyderabad.

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Abstract

The drug development undergoes series of clinical trials in which there is a possibility of obtaining false positive results for higher mammals. In order to reduce the number of experiments involving animals, there is an urgent need to introduce an intermediate stage. Specifically, low cost cell culture models in a controlled environment is gaining importance in toxicity studies. Although there are many cell culture model in petri dishes, they consume higher volume of media and reagents. In this scenario, 3D printing is used for manufacturing devices and rapid prototyping rather than traditional techniques including soft lithography. Although there is some effort in the direction of fabrication of devices compatible to cell culture through 3D printing, to the best of our knowledge, there is no co-culture device that can be used for culturing multiple cells, imaging the cell morphologies and perfuse multiple drugs. Here we present a fabrication protocol for a glass bottom PDMS based coculture device that contain two interconnected chambers and four inlets for mixing drugs through passive mixing regions. First we compared the printing of negative template using fused deposition modeling and stereo lithography. Next we evaluate the printing error and fidelity of printing for the two printers. Specifically, we performed a series of experiments to select the dimension of the channel for which the device can be printed reproducible with less than 5% error. Measurement of dimension was performed through panorama imaging using laser scanning confocal microscopy. Moreover, characterization of roughness of the printed surface was performed using an optical profiler. Almost ten versions were fabricated to achieve the optimized device dimension and the proper bonding protocols. The salient features of the proposed fabrication are the fabrication of the negative template embedded in a box, bonding of the PDMS structure with the glass slide using the thin coat of PDMS as glue and compatibility to Nikon phase contrast microscope for imaging. Here we also show that the proposed bonding technique gives rise to a leak proof structure that is compatible to cell growth for longer period of time. Specifically, we show a proof of concept that the neuronal cells can be maintained in the device for 48 hours. Such co-culture device can be used in future for culturing vii multiple cell types including neuron and glial cells. It can also be used for testing multiple drugs through perfusion of liquid mixture containing imaging buffer and drugs. Some of the specific advantages of the proposed method based on 3D printing include the reproducibility in device dimensions, low cost set up and rapid prototyping of complex designs

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