To evaluate whether a purified human platelet lysate (HPL) and platelet extracellular vesicles (P-EV) rich in platelet growth factors can enhance proliferation and repair of corneal endothelial cells (CECs) in vitro
Methods
HPL and P-EV were prepared from human clinical-grade platelet concentrates (PC). Platelets were isolated by centrifugation and lysed by three freeze-thaw cycles to release the growth factors. The lysate was heat-treated, centrifuged, and the supernatant (HPL) recovered. P-EV were isolated by sequential centrifugation of the plasma compartment of PC. HPLs and P-EV were characterized for protein and growth factors content and platelet surface markers. Viability of two kinds of CECs: B4G12 and BCE C/D-1b cells exposed to HPL and P-EV was evaluated by CCK-8 proliferation assay and morphology was observed by microscopic observation after 24 and 48 hours of culture. CEC migration was assessed using a wound healing scratch assay after 0, 6, 12 and 24 hours of treatment.
Results
HPL and P-EV contained growth factors (TGFβ, EGF, VEGF, IGF, PDGF-BB) known to be beneficial for CEC growth. Treatment of B4G12 and BCE C/D-1b cells with 5% HPL and 60ug/ml P-EV maintained cell growth and morphology, and increased cell viability compared to untreated control (medium with 5% FBS and 10ng/ml bFGF). Furthermore, HPL and P-EV enhanced the migration of B4G12 and BCE C/D-1b cells subjected to the scratch assay compared to control.
Conclusion
HPL and P-EV made from clinical-grade PC enhance the proliferation and migration of CECs, and could thus be considered in a repair strategy protocol of the damaged corneal endothelium.
Conflict of interest
No
Author 1
Last name
WIDYANINGRUM
Initials of first name(s)
R
City
Taipei
Country
Taiwan
Author 2
Last name
Wang
Initials of first name(s)
T-J
City
Taipei
Country
Taiwan
Author 3
Last name
Burnouf
Initials of first name(s)
T
City
Taipei
Country
Taiwan
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