Platelet-Rich Plasma Can Replace Fetal Bovine Serum in Human Meniscus Cell Cultures

Abstract

Concerns over fetal bovine serum (FBS) limit the clinical application of cultured tissue-engineered constructs. Therefore, we investigated if platelet-rich plasma (PRP) can fully replace FBS for meniscus tissue engineering purposes. Human PRP and platelet-poor plasma (PPP) were isolated from three healthy adult donors. Human meniscal fibrochondrocytes (MFCs) were isolated from resected tissue after a partial meniscectomy on a young patient. Passage-4 MFCs were cultured in monolayer for 24 h, and 3 and 7 days. Six different culture media were used containing different amounts of either PRP or PPP and compared to a medium containing 10% FBS. dsDNA was quantified, and gene expression levels of collagen types I and II and aggrecan were measured at different time points with quantitative polymerase chain reaction in the cultured MFCs. After 7 days, the dsDNA quantity was significantly higher in MFCs cultured in 10% and 20% PRP compared to the other PRP and PPP conditions, but equal to 10% FBS. Collagen type I expression was lower in MFCs cultured with medium containing 5% PRP, 10% and 20% PPP compared to FBS. When medium with 10% PRP or 20% PRP was used, expressions were not significantly different from medium containing 10% FBS. Collagen type II expression was absent in all medium conditions. Aggrecan expression did not show differences between the different media used. However, after 7 days a higher aggrecan expression was measured in most culture conditions, except for 5% PRP, which was similar compared to FBS. Statistical significance was found between donors at various time points in DNA quantification and gene expression, but the same donors were not statistically different in all conditions. At 7 days cell cultured with 10% PRP and 20% PRP showed a higher density, with large areas of clusters, compared to other conditions. In an MFC culture medium, FBS can be replaced by 10% PRP or 20% PRP without altering proliferation and gene expression of human MFCs.

Platelet Rich Concentrate Promotes Early Cellular Proliferation and Multiple Lineage Differentiation of Human Mesenchymal Stromal CellsIn Vitro

“…The use of activated PRC provided no clear advantage over nonactivated PRC on the proliferation of cells. Thus, the findings of this study support the notion that, in view of its additive proliferative effect, PRC could potentially substitute the commercially available FBS or recombinant growth factors that are normally used for MSC expansion in vitro , as suggested by previous studies [ 16 , 23 ].…”

A novel biphasic scaffold supports meniscal tissue repair in ex vivo and in vivo models

“…It has been shown that MFCs are mobilised by signals found in fibrin clots [33] and that platelet derived growth factor (PDGF) and endothelin-1 promote migration of meniscal cells in vitro [30] . In this study we have demonstrated that the addition of PRP significantly enhanced the migration of the MFC into the scaffolds and increased the deposition of ECM within the scaffolds, as has been reported in other studies [12,34] . PRP contains both fibrin and a number of growth factors, including PDGF, any of which could be acting alone or in synergy to enhance MFC migration.…”

Platelet-rich plasma: why intra-articular? A systematic review of preclinical studies and clinical evidence on PRP for joint degeneration

“…PRP up-regulated the viability of meniscal cells in a dose-dependent manner, as well as the mRNA expression of biglycan and decorin. Gonzales et al [ 23 ] investigated whether PRP might fully replace FBS for cultured tissue engineering constructs. The study results showed that PRP presents the same positive effect as FBS for meniscal cell culture and showed that dosage is an important aspect of the induced effect: 10 and 20 % PRP increased proliferation rate and influenced more type I collagen and aggrecan expression at day 7 of culture with respect to 5 % PRP.…”