The Establishment Of Green Fluorescent Clones Of A549 Human Lung Carcinoma Cells

Hannah Addis

School Name

Governor's School for Science and Math

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Mentor

Mentor: Dr. Wei; Department of Biology, Clemson University

Abstract

A549 human lung carcinoma cells are widely used in animal model studies for human anticancer therapy development. The detection of A549 tumor cells in animals requires complicated reagents and procedures. The goal of this project is to create clones of A549 cells that express green fluorescent protein (GFP) so that the tumor cells can be easily recognized in vivo. A549 cells were cultured and plated in a six-well plate. The cells were then transfected with plasmid pcDNA3.1-GFP/zeo+ via Lipofectamine 2000. After transfection, a selection media containing zeocin was used to select transfected cells. The fluorescent imaging results demonstrated that A549 cells were successfully transfected with GFP. Many colonies resistant to zeocin were established but did not express GFP. Further attempts to obtain stable colonies will be explored.

Location

Owens 201

Start Date

4-16-2016 8:45 AM

COinS
 
Apr 16th, 8:45 AM

The Establishment Of Green Fluorescent Clones Of A549 Human Lung Carcinoma Cells

Owens 201

A549 human lung carcinoma cells are widely used in animal model studies for human anticancer therapy development. The detection of A549 tumor cells in animals requires complicated reagents and procedures. The goal of this project is to create clones of A549 cells that express green fluorescent protein (GFP) so that the tumor cells can be easily recognized in vivo. A549 cells were cultured and plated in a six-well plate. The cells were then transfected with plasmid pcDNA3.1-GFP/zeo+ via Lipofectamine 2000. After transfection, a selection media containing zeocin was used to select transfected cells. The fluorescent imaging results demonstrated that A549 cells were successfully transfected with GFP. Many colonies resistant to zeocin were established but did not express GFP. Further attempts to obtain stable colonies will be explored.