title: Expression of green fluorescent protein in human foreskin fibroblasts for use in 2D and 3D culture models.
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.
authors.affiliation: Departments of Surgery, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Chao
authors.forename: Tiffany
authors.initials: T
authors.lastname: Peña
authors.forename: Dean G
authors.initials: DG
authors.lastname: Heimann
authors.forename: Chris
authors.initials: C
authors.lastname: Hansen
authors.forename: David A
authors.initials: DA
authors.lastname: Doyle
authors.forename: Ujwal R
authors.initials: UR
authors.lastname: Yanala
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Guenther
authors.forename: Mark A
authors.initials: MA
authors.lastname: Carlson
mesh_terms: D018929:Cell Culture Techniques; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D052816:Foreskin / Q000166:cytology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D049408:Luminescent Agents / Q000378:metabolism*; D008297:Male
chemical_list: D049408:Luminescent Agents; D049452:Green Fluorescent Proteins; D003094:Collagen