Date of Completion
Honors College Thesis
Department of Pulmonary Medicine
Daniel Weiss, MD, PhD
Janet Murray, PhD
lung therapy, cell culture, regenerative medicine, bioengineering
The prevalence of lung diseases such as chronic obstructive pulmonary disease (COPD) is on the rise and with it the demand for donor lungs for transplantation in end-stage disease patients (Wagner et al., 2013). This demand continually exhausts the available donor lung supply, therefore, novel alternatives for lung transplantation are being investigated. One of these potential solutions is the creation of a bioengineered synthetic lung for use in transplantation. A successful methodology would result in an independence from the limited supply of donor lungs as well as a solution to immune rejection by the transplant recipient, as cells from the recipient could be used to cellularize the engineered lung and minimize rejection. A proposed protocol for the synthesis of bioengineered lungs involves the use of a lung scaffold, obtained by removing all cellular material from a pre-existing lung, leaving on the extracellular matrix proteins, and seeding it with patient cells to grow a new, viable lung (Uriarte et al., 2018). However, one problem that currently hinders progress is the ability to seed multiple cell lines on a single scaffold. Currently a single cell line can be seeded, however this is not representative of a completely functioning lung possessing over 40 types of cells. Therefore, this study focused on developing a preliminary protocol on how to co-seed multiple cell lines, traditionally cultured in different growth media, on a single scaffold. In this study, cell culture was conducted with human lung fibroblast cells (HLFs) and human bronchial epithelial cells (HBEs). The two cell lines were initially grown and observed in their own respective media to determine proportional growth rates. They were then seeded individually and as co-seeds in plates varying in media components and in cell counts. It was found that a co-seed of HLF and HBE cells is possible over a seven-day period in a media composed of 50% HBE and 50% HLF media. Growth in this trial followed an exponential trend, and the cells were interspersed indicating that they were cohabitating in a healthy manner.
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Nuckols, Christopher R., "Optimizing Cell Culture Media for Combined Use for Ex Vivo Lung Bioengineering" (2020). UVM Honors College Senior Theses. 359.