GMP-grade platelet lysate enhances proliferation and migration of tenon fibroblasts

Abstract

Tenon's fibroblasts (TFs), widely employed as in vitro model for many ophthalmological studies, are routinely cultured with FBS. Platelet Lysate (PL), a hemoderivate enriched with growth factors and cytokines has been largely tested in several clinical applications and as substitute of FBS in culture. Here, we investigate whether PL can exert biological effects on TF populations similarly to other cell types. Results show that PL significantly enhances cell proliferation and migration vs. FBS, without influencing cell size/granularity. Upregulation of EGF, VEGF, KDR, MMP2-9, FAK mRNA levels also occurs and phosphorylation of AKT but not of ERK1/2 is significantly enhanced. The inhibition of the PI3kinase/AKT pathway with the specific inhibitor wortmannin, decreases PL-induced cell migration but not proliferation. Condition supernatants containing PL show increased bioavailability of Nitric Oxide and reduced levels of 8-Iso-PGF2-alpha, correlating with cell proliferation and migration. Pro-angiogenic/inflammatory soluble factors (GRO, Angiogenin, EGF, I-309, PARC) are exclusively or greater expressed in media containing PL than FBS. GMP-grade PL preparations positively influence in vitro biological effects of TFs representing a suitable and safer alternative to FBS.

Remote Adipose Tissue-Derived Stromal Cells of Patients with Lung Adenocarcinoma Generate a Similar Malignant Microenvironment of the Lung Stromal Counterpart

“…FACS analysis was performed to investigate the percentage of apoptotic cells after stimulation with AMSC-derived conditioned media as previously reported [ 30 , 31 ]. Briefly, semiconfluent cultures were harvested with Accutase (Sigma-Aldrich) and stained for 30 minutes with 5 μ l AnnexinV-FITC antibody (Invitrogen Cat.…”

“…The evaluation of the different microenvironments was performed on collected supernatant obtained from both A- and LMSC in patients with lung adenocarcinoma and hemartochondroma. C-Series Human Cytokine Antibody Array C5 (RayBiotech, Inc) was used for simultaneous semiquantitative detection of 80 multiple cytokines/growth factors as previously described [ 31 ]. Briefly, an equal volume of collected undiluted supernatants was incubated by gentle shaking overnight at 4°C on the membrane of the kit C-Series Human Cytokine Antibody Array C5.…”