Expansion of Human Dermal Papilla Cells for Clinical Applications Using Human Platelet-rich Plasma as a Substitute...

Cell expansion for clinical cell culture often involves the use of fetal bovine serum (FBS). Recently, human platelet-rich plasma (hPRP) was suggested as an FBS substitute, to minimize the risk of transmission of various pathogens and immunogenic reactions. However, whether hPRP can replace FBS for the expansion of human dermal papilla cells (hDPCs) remains controversial.

To assess the potential of hPRP as a substitute for FBS in hDPC expansion, we investigated the effect of hPRP on hDPC expansion.

We first standardized the method of hPRP preparation, and then investigated the effect of hPRP on the proliferation and hair-inductive capacity of cultured hDPCs by comparing with that of FBS. To prepare hPRP, we used heparin as the anticoagulant, with purification by single-spin centrifugation, and freeze-thaw hPRP activation.

Compared with an FBS-supplemented medium, hPRP supplementation significantly shortened the population doubling time of cultured hDPCs, and increased the ratio of bromodeoxyuridine-labeled proliferating cells. Expression of the alkaline phosphatase gene related to hair-inductive capacity was enhanced in hDPCs cultured in an hPRP-supplemented medium. In vivo hair follicle reconstitution assay confirmed that hPRP supplementation enhanced the hair-inductive capacity of hDPCs compared with an FBS-supplemented medium, based on the increased number of regenerated hair follicles and their maturity. Overall, compared with FBS, hPRP enhanced the proliferation of hDPCs, while preserving their hair-inductive capacity.

The results show that hPRP can be used to replace FBS for hDPC expansion. These findings inform the development of an effective clinically applicable hair regeneration therapy.