1. Expression of green fluorescent protein in human foreskin fibroblasts for use in 2D and 3D culture models.
- Author
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Chao, Jie, Peña, Tiffany, Heimann, Dean G., Hansen, Chris, Doyle, David A., Yanala, Ujwal R., Guenther, Timothy M., and Carlson, Mark A.
- Subjects
FIBROBLASTS ,PROTEIN metabolism ,ANALYSIS of variance ,GENE expression ,IMMUNOHISTOCHEMISTRY ,MATHEMATICAL models ,RESEARCH methodology ,RESEARCH funding ,T-test (Statistics) ,TISSUE culture ,THEORY ,DESCRIPTIVE statistics ,IN vitro studies ,PHYSIOLOGY - Abstract
The availability of fibroblasts that express green fluorescent protein ( GFP) would be of interest for the monitoring of cell growth, migration, contraction, and other processes within the fibroblast-populated collagen matrix and other culture systems. A plasmid lentiviral vector-GFP (pLV-GFP) was utilized for gene delivery to produce primary human foreskin fibroblasts ( HFFs) that stably express GFP. Cell morphology, cell migration, and collagen contraction were compared between nontransduced HFFs and transduced GFP- HFFs; no differences were observed. Immunocytochemical staining showed no differences in cell morphology between nontransduced and GFP- HFFs in both two-dimensional and three-dimensional culture systems. Furthermore, there was no significant difference in cellular population growth within the collagen matrix populated with nontransduced vs. GFP- HFFs. Within the limits of our assays, we conclude that transduction of GFP into HFFs did not alter the observed properties of HFFs compared with nontransduced fibroblasts. The GFP- HFFs may represent a new tool for the convenient monitoring of living primary fibroblast processes in two-dimensional or three-dimensional culture. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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