A Study on the Effect of Media Volume Manipulations on Global HUVEC Colony Proliferation Within a Microfluidic Channel

Open Access
Author:
Poudyal, Ashutosh
Graduate Program:
Bioengineering
Degree:
Master of Science
Document Type:
Master Thesis
Date of Defense:
April 10, 2014
Committee Members:
  • Siyang Zheng, Thesis Advisor
  • Sheereen Majdzarringhalamaraghy, Thesis Advisor
  • William O Hancock, Thesis Advisor
Keywords:
  • HUVEC
  • microfluidic
  • cell culture
Abstract:
Microfluidic-based cell culture is a rapidly developing subject used in studying behavior and external influences on cells on a micro-scale. A majority of modern techniques utilize a perfusion-based culture model where fresh media is continuously passed over growing cells in order to keep them replenished. However, such techniques require intricate setups that have to incorporate syringe pumps, the growth platform, and a culture incubator. To add to the complexity, sterility has to be maintained so as not to contaminate the cells. Unlike perfusion-based techniques, this work demonstrates a novel method to culture cells utilizing a culture media reservoir and nutrient diffusion. Two micropipette tips fixed on the inlet/outlet ends of a Polydimethylsiloxane (PDMS) microfluidic culture channel are filled with media that passively diffuses throughout the channel to sustain the cells. Optimal times and techniques to replenish the media reservoir were determined through 3D computer simulation and wet bench experimentation. Cell growth on a glass, PDMS, and a Matrigel platform were explored. Ultimately, it was shown that replenishing the culture media within the reservoirs in increments of 5μL allowed for a confluent Human umbilical vein endothelial (HUVE) cell colony to grow in 32 hours of culture. Replenishing the media in 20μL or 50μL increments did not foster global colony growth. Further computational image analysis of cell culture images showed the 5μL culture technique led to roughly two times greater cell colony coverage throughout the growth channel.