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

Abstract

Platelet rich concentrate (PRC) is a natural adjuvant that aids in human mesenchymal stromal cell (hMSC) proliferation in vitro; however, its role requires further exploration. This study was conducted to determine the optimal concentration of PRC required for achieving the maximal proliferation, and the need for activating the platelets to achieve this effect, and if PRC could independently induce early differentiation of hMSC. The gene expression of markers for osteocytes (ALP, RUNX2), chondrocytes (SOX9, COL2A1), and adipocytes (PPAR-γ) was determined at each time point in hMSC treated with 15% activated and nonactivated PRC since maximal proliferative effect was achieved at this concentration. The isolated PRC had approximately fourfold higher platelet count than whole blood. There was no significant difference in hMSC proliferation between the activated and nonactivated PRC. Only RUNX2 and SOX9 genes were upregulated throughout the 8 days. However, protein expression study showed formation of oil globules from day 4, significant increase in ALP at days 6 and 8 (P ≤ 0.05), and increased glycosaminoglycan levels at all time points (P < 0.05), suggesting the early differentiation of hMSC into osteogenic and adipogenic lineages. This study demonstrates that the use of PRC increased hMSC proliferation and induced early differentiation of hMSC into multiple mesenchymal lineages, without preactivation or addition of differentiation medium.

Platelet-rich concentrate in serum free medium enhances osteogenic differentiation of bone marrow-derived human mesenchymal stromal cells

“…In our preparation, the concentration of PDGF-AA, PDGF-AB, PDGF-BB and TGF- β 1 was found to be 50.32 ng/mL, 9.91 ng/mL, 8.32 ng/mL and 15.17 ng/mL respectively. These results were previously reported elsewhere ( Shani et al, 2014 ).…”

“…Twenty-five milliliters of blood from each subject was collected in ACD tubes. Platelets were prepared using double centrifugation method as mentioned previously, with slight modifications ( Cho et al, 2011 ; Lee et al, 2012 ; Shani et al, 2014 ). In brief, the first centrifugation was at 200× g for 10 min, which separated the red blood cells, plasma and buffy coat that contains the platelets and the white blood cells.…”

A Proliferation and surface marker characterization of adipose stem cells after culture in various processed outdated platelet lysate containing media

“…Another study by Shani et al 23 performed comparative study between activated platelet using 10% CaCl 2 and human thrombin (1:10 [v/v]) and non-activated platelet, and reported that cell proliferations reached the peak of proliferation at 8 days after culture of human bone marrowmesenchymal stem cells with viable cell numbers of > 60.000 cells with a seeding number of > 10.000 cells/well. They also stated that there was no difference in cell proliferation between activated and nonactivated platelet.…”

“…However, in our study, at day-7, viable cell number reach > 500.000 cells/well on F1, F2, and F3 medium. Shani et al 23 added medium only once at the beginning of culture without medium change through all experiment. Without frequent medium change, the culture might lead to generation of metabolite/waste products, mainly lactate and ammonia that lead to decrease in cell growth rate.…”

Mesenchymal stromal cells for cartilage repair in osteoarthritis

“…Also human MSCs have been isolated from several tissues including peripheral blood 13,14 . Regardless of their origin they have the capacity to differentiate into many cell types depending on the stimulus 15 . One of the important properties of MSCs is to retain immunomodulatory activity due to lack of human leucocyte antigen (HLA) class II expression 16,17 .…”