Brief communication: Effects of conditioned media from human platelet lysate cultured MSC on osteogenic cell differentiation in vitro

Abstract

Culturing mesenchymal stromal cells (MSC) in human platelet lysate (HPL) supplemented media can enhance their osteogenic differentiation potential. The objective of this study was to test the hypothesis that conditioned media (CM) derived from HPL-cultured MSC also have pro-osteogenic effects. Pooled CM was prepared from HPL-cultured human bone marrow MSC (BMSC) of multiple donors and applied on BMSC of different donors (than those used for CM preparation), with or without additional supplementation [HPL, fetal bovine serum (FBS)] and osteogenic stimulation. At various time-points, cell proliferation, alkaline phosphatase (ALP) activity, osteogenic gene expression and in vitro mineralization were assessed. BMSC in standard unstimulated growth media served as controls. After 3–7 days, CM alone did not promote BMSC proliferation or ALP activity; supplementation of CM with HPL slightly improved these effects. After 2 and 7 days, CM alone, but not CM supplemented with HPL, promoted osteogenic gene expression. After 14 days, only CM supplemented with FBS and osteogenic stimulants supported in vitro BMSC mineralization; CM alone and CM supplemented with HPL did not support mineralization, regardless of osteogenic stimulation. In summary, CM from HPL-cultured BMSC promoted osteogenic gene expression but not in vitro mineralization in allogeneic BMSC even when supplemented with HPL and/or osteogenic stimulants. Future studies should investigate the role and relevance of supplementation and osteogenic induction in in vitro assays using CM from MSC.

The use of mesenchymal stromal cell secretome to enhance guided bone regeneration in comparison with leukocyte and platelet-rich fibrin

“…We have previously reported the pro-osteogenic effects of MSC-CM on human BMSC (40). Consistently, in the present study, more osteogenesis related genes, namely RUNX2, COL1A2, SPP1/OPN and BGLAP/OCN, were upregulated in BMSC exposed to MSC-CM vs. PRF-CM after 48 h in monolayer culture (Fig.…”

“…MSC-CM was prepared from BMSC of three donors as previously described (40). Brie y, p1 and p2 BMSC were expanded in GM until 70-80% con uency under standard incubation.…”

The use of mesenchymal stromal cell secretome to enhance guided bone regeneration in comparison with leukocyte and platelet‐rich fibrin

“…MSC‐CM was prepared from BMSC of three donors as previously described (Shanbhag et al., 2022). Briefly, p1 and p2 BMSC were expanded in growth media until 70%–80% confluency under standard incubation.…”

Runx2 overexpression promotes bone repair of osteonecrosis of the femoral head (ONFH)

“…However, Simple MSCs transplantation have some shortcomings, such as a large number of cells are needed, multiple transplants are needed, long time to repair is needed and unstable differentiation ability without induction. Some methods have been used to induce MSCs differentiate into osteoblasts, These methods mainly include drugs, physical stimulation, exosomes, gene transfection and platelet lysate and so on [15][16][17][18][19][20][21][22].…”