Phenotypic study of human gingival fibroblasts in a medium enriched with platelet lysate.
BACKGROUND
The modulation abilities of gingival fibroblasts open new therapeutic strategies for the treatment of vascular diseases (e.g., aneurism) and irradiation burns. Culture media are classically supplemented with animal sera to provide nutriments. Unfortunately, because of their potential for interspecies transmission of microorganisms, these media are not used for cells destined for human transplantation. This preliminary phenotypic study aims to test a serum-free (SF) culture medium for human gingival fibroblasts (hGF) supplemented with human platelet lysates (PLs) for rapid cell expansion.


METHODS
An SF medium was first elaborated to compete with hGF proliferation in a reference medium containing 10% fetal bovine serum (BSmedium). Adhesion, proliferation, and doubling kinetics were run in the presence of PLs (SF+PL). Cytoskeletal proteins were analyzed and chromosomal abnormalities were evaluated by karyotype analyses. The SF+PL influence on secretion of molecules implied in tissue remodeling (i.e., matrix metalloproteinases [MMPs], their tissue inhibitors [TIMPs], and several growth factors) was studied.


RESULTS
SF+PL increased the proliferation rate 1.5-fold in a week compared to BSmedium. Cytoskeleton protein expression was similar in BSmedium and in SF+PL. Chromosomal abnormalities were rare in SF+PL. MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, TIMP-1, and the growth factors interleukin-1β and -4 and transforming growth factor-β1 secretions were stable during the experiment. TIMP-2 and interleukin-6 were slightly decreased in SF+PL compared to BSmedium.


CONCLUSIONS
While waiting confirmation from a proteomic approach, this SF culture medium could allow a secured faster hGF proliferation adapted for human cell transplant therapy.
Institute for Health and Medical Research - Unit 849, Paris Descartes University, Health Services-Paris Hospitals, Paris, France. adrien.naveau@laposte.net
Adrien
A
Naveau
Jean-Jacques
JJ
Lataillade
Benjamin Philippe
BP
Fournier
Ludovic
L
Couty
Marie
M
Prat
François Côme
FC
Ferre
Muriel
M
Gourven
Eric
E
Durand
Bernard
B
Coulomb
Antoine
A
Lafont
Bruno
B
Gogly
D001792:Blood Platelets / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D016895:Culture Media, Serum-Free; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D005881:Gingiva / Q000166:cytology; D006801:Humans; D007424:Intracellular Fluid / Q000502:physiology; D010641:Phenotype; D010865:Pilot Projects
D016895:Culture Media, Serum-Free