Primary cultured fibroblasts derived from patients with chronic wounds: a methodology to produce human cell lines and test putative growth factor therapy such as GMCSF.
BACKGROUND
Multiple physiologic impairments are responsible for chronic wounds. A cell line grown which retains its phenotype from patient wounds would provide means of testing new therapies. Clinical information on patients from whom cells were grown can provide insights into mechanisms of specific disease such as diabetes or biological processes such as aging. The objective of this study was 1) To culture human cells derived from patients with chronic wounds and to test the effects of putative therapies, Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) on these cells. 2) To describe a methodology to create fibroblast cell lines from patients with chronic wounds.


METHODS
Patient biopsies were obtained from 3 distinct locations on venous ulcers. Fibroblasts derived from different wound locations were tested for their migration capacities without stimulators and in response to GM-CSF. Another portion of the patient biopsy was used to develop primary fibroblast cultures after rigorous passage and antimicrobial testing.


RESULTS
Fibroblasts from the non-healing edge had almost no migration capacity, wound base fibroblasts were intermediate, and fibroblasts derived from the healing edge had a capacity to migrate similar to healthy, normal, primary dermal fibroblasts. Non-healing edge fibroblasts did not respond to GM-CSF. Six fibroblast cell lines are currently available at the National Institute on Aging (NIA) Cell Repository.


CONCLUSIONS
We conclude that primary cells from chronic ulcers can be established in culture and that they maintain their in vivo phenotype. These cells can be utilized for evaluating the effects of wound healing stimulators in vitro.
Brem
Harold
H
Department of Surgery, Division of Wound Healing & Regenerative Medicine, New York University School of Medicine, New York, NY, USA. Harold.Brem@nyumc.org
Golinko
Michael S
MS
Stojadinovic
Olivera
O
Kodra
Arber
A
Diegelmann
Robert F
RF
Vukelic
Sasa
S
Entero
Hyacinth
H
Coppock
Donald L
DL
Tomic-Canic
Marjana
M
D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014647:Varicose Ulcer / Q000473:pathology; D014945:Wound Healing / Q000187:drug effects* / Q000502:physiology
D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor