Shanbhag Siddharth, Al-Sharabi Niyaz, Mohamed-Ahmed Samih, Gruber Reinhard, Kristoffersen Einar K, Mustafa Kamal
Department of Immunology and Transfusion Medicine, Haukeland University Hospital, Bergen, Norway.
Center for Translational Oral Research, Department of Clinical Dentistry, Faculty of Medicine, University of Bergen, Bergen, Norway.
Front Bioeng Biotechnol. 2022 Sep 29;10:969275. doi: 10.3389/fbioe.2022.969275. eCollection 2022.
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 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 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 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 assays using CM from MSC.
在添加人血小板裂解物（HPL）的培养基中培养间充质基质细胞（MSC）可以增强其成骨分化潜能。本研究的目的是检验以下假设：源自HPL培养的MSC的条件培养基（CM）也具有促骨生成作用。从多个供体的HPL培养的人骨髓MSC（BMSC）中制备混合CM，并将其应用于不同供体的BMSC（不同于用于制备CM的供体），同时添加或不添加其他成分[HPL、胎牛血清（FBS）]以及进行成骨刺激。在不同时间点，评估细胞增殖、碱性磷酸酶（ALP）活性、成骨基因表达和矿化情况。在标准未刺激生长培养基中的BMSC作为对照。3 - 7天后，单独的CM不促进BMSC增殖或ALP活性；用HPL补充CM可略微改善这些作用。2天和7天后，单独的CM促进成骨基因表达，但用HPL补充的CM则没有。14天后，只有补充了FBS和成骨刺激剂的CM支持BMSC矿化；单独的CM以及用HPL补充的CM，无论是否有成骨刺激，均不支持矿化。总之，来自HPL培养的BMSC的CM即使补充了HPL和/或成骨刺激剂，也能促进同种异体BMSC的成骨基因表达，但不支持矿化。未来的研究应调查在使用MSC的CM的实验中补充成分和成骨诱导的作用及相关性。
