Presentation Title

Developing a Protocol to Co-Seed Multiple Cell Lines in the same Media

Presenter's Name(s)

Christopher NuckolsFollow

Project Collaborators

Daniel Weiss (PI)

Abstract

There is a high demand for donor lungs for lung transplantation, as the prevalence of lung diseases is increasing and there are limited donor organs to meet the demand 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 donor lungs, as well as a solution to host rejection, as host stem cells could be used to cellularize the synthetic lung. A proposed protocol for the synthesis 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. A problem that currently hinders progress is the seeding of multiple cell lines on a single scaffold. Currently, a single cell line can be seeded, however, as the lung has over 40 types of cells, this is not representative of a completely functioning lung. This study will focus on developing a preliminary protocol on how to co-seed multiple cell lines on a single scaffold at once. In this study, cell culture was conducted with human lung fibroblastic cells and human bronchial epithelial cells. The two cell lines were initially grow and observed in their own respective media to determine proportional growth rates. They were then seeded individual and as co-seeds in plates varying in media concoctions and cellular concentrations. 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 instead of segregated indicating that they were cohabitating in a healthy manner.

Primary Faculty Mentor Name

Daniel Weiss

Status

Undergraduate

Student College

College of Agriculture and Life Sciences

Program/Major

Microbiology

Primary Research Category

Biological Sciences

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Developing a Protocol to Co-Seed Multiple Cell Lines in the same Media

There is a high demand for donor lungs for lung transplantation, as the prevalence of lung diseases is increasing and there are limited donor organs to meet the demand 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 donor lungs, as well as a solution to host rejection, as host stem cells could be used to cellularize the synthetic lung. A proposed protocol for the synthesis 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. A problem that currently hinders progress is the seeding of multiple cell lines on a single scaffold. Currently, a single cell line can be seeded, however, as the lung has over 40 types of cells, this is not representative of a completely functioning lung. This study will focus on developing a preliminary protocol on how to co-seed multiple cell lines on a single scaffold at once. In this study, cell culture was conducted with human lung fibroblastic cells and human bronchial epithelial cells. The two cell lines were initially grow and observed in their own respective media to determine proportional growth rates. They were then seeded individual and as co-seeds in plates varying in media concoctions and cellular concentrations. 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 instead of segregated indicating that they were cohabitating in a healthy manner.