Platelet-rich plasma enhances the proliferation of human adipose stem cells through multiple signaling pathways

Stem Cell Research & Therapy, Apr 2018

Background Platelet-rich plasma (PRP) is an autologous blood product that contains a high concentration of several growth factors. Platelet-derived growth factor (PDGF)-BB is a potential mitogen for human adipose-derived stem cells (hASCs). PRP stimulates proliferation of hASCs; however, the signaling pathways activated by PRP remain unclear. Methods hASCs were cultured with or without PRP or PDGF-BB, and proliferation was assessed. hASCs were also treated with PRP or PDGF-BB with or without imatinib, which is a PDGF receptor tyrosine kinase inhibitor, or sorafenib, which is a multikinase inhibitor. Inhibition of cell proliferation was examined using anti-PDGF antibody (Abcam, Cambridge, UK), by cell counting. We assessed the effects of inhibitors of various protein kinases such as ERK1/2, JNK, p38, and Akt on the proliferation of hASCs. Results The proliferation was remarkably promoted in cells treated with either 1% PRP or 10 ng/ml PDGF-BB, and both imatinib and sorafenib inhibited this proliferation. Anti-PDGF antibody (0.5 and 2 μg/ml) significantly decreased the proliferation of hASCs compared with control. PRP-mediated hASC proliferation was blocked by inhibitors of ERK1/2, Akt, and JNK, but not by an inhibitor of p38. Conclusions PRP promotes hASC proliferation, and PDGF-BB in PRP plays a major role in inducing the proliferation of hASCs. PRP promotes hASC proliferation via ERK1/2, PI3K/Akt, and JNK signaling pathways.

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Platelet-rich plasma enhances the proliferation of human adipose stem cells through multiple signaling pathways

Lai et al. Stem Cell Research & Therapy Platelet-rich plasma enhances the proliferation of human adipose stem cells through multiple signaling pathways Fangyuan Lai 0 Natsuko Kakudo 0 Naoki Morimoto 0 Shigeru Taketani 2 Tomoya Hara 0 1 Takeshi Ogawa 0 Kenji Kusumoto 0 0 Department of Plastic and Reconstructive Surgery, Kansai Medical University , 2-5-1 Shin-machi, Hirakata, Osaka 573-1010 , Japan 1 Department of Oral Implantology, Osaka Dental University , Osaka 573-1121 , Japan 2 Department of Microbiology, Kansai Medical University , Osaka 573-1010 , Japan Background: Platelet-rich plasma (PRP) is an autologous blood product that contains a high concentration of several growth factors. Platelet-derived growth factor (PDGF)-BB is a potential mitogen for human adipose-derived stem cells (hASCs). PRP stimulates proliferation of hASCs; however, the signaling pathways activated by PRP remain unclear. Methods: hASCs were cultured with or without PRP or PDGF-BB, and proliferation was assessed. hASCs were also treated with PRP or PDGF-BB with or without imatinib, which is a PDGF receptor tyrosine kinase inhibitor, or sorafenib, which is a multikinase inhibitor. Inhibition of cell proliferation was examined using anti-PDGF antibody (Abcam, Cambridge, UK), by cell counting. We assessed the effects of inhibitors of various protein kinases such as ERK1/2, JNK, p38, and Akt on the proliferation of hASCs. Results: The proliferation was remarkably promoted in cells treated with either 1% PRP or 10 ng/ml PDGF-BB, and both imatinib and sorafenib inhibited this proliferation. Anti-PDGF antibody (0.5 and 2 μg/ml) significantly decreased the proliferation of hASCs compared with control. PRP-mediated hASC proliferation was blocked by inhibitors of ERK1/2, Akt, and JNK, but not by an inhibitor of p38. Conclusions: PRP promotes hASC proliferation, and PDGF-BB in PRP plays a major role in inducing the proliferation of hASCs. PRP promotes hASC proliferation via ERK1/2, PI3K/Akt, and JNK signaling pathways. Background Human adipose-derived stem cells (hASCs) were first isolated from human adipose tissue and identified by Zuk et al. in 2001 [ 1 ]. These cells can differentiate toward multiple lineages, such as osteogenic [ 2 ], chondrogenic [ 3 ], adipogenic [ 4 ], cardiac [ 5 ], epidermal [ 6 ], and neurogenic [ 7 ] lineages. hASCs are used widely in the field of regenerative medicine, including to promote bone regeneration [ 2 ], tooth and periodontal regeneration [ 8 ], cartilage regeneration [ 9 ], wound healing [ 6, 10 ], and nerve regeneration to cure Parkinson’s disease [11], as well as to suppress aging [ 10 ]. Due to the advantages of the autologous source of these cells and their relative abundance and ease of isolation, hASCs have also been widely used in the fields of plastic surgery and regenerative medicine [ 12 ]. However, the proliferation and differentiation capacities of hASCs decrease with age [ 13, 14 ], body mass index [14], diabetes mellitus [ 12, 15 ], radiation exposure [16], and tamoxifen treatment [ 17 ]. hASCs account for about 16–30% of the stromal vascular fraction [ 18 ]. To obtain a sufficient amount of cells for therapeutic purposes, in-vitro proliferation of the cells is required. Fetal bovine serum (FBS) is widely used for this purpose in multiple types of cells in vitro. However, due to the risk of heterologous immunization and zoonosis, FBS has limited clinical use. Platelet-rich plasma (PRP) is a blood portion that is enriched with platelets [ 19 ]. Upon activation, platelets in PRP release granules containing molecules including growth factors and regulatory proteins, such as plateletderived growth factor (PDGF), epidermal growth factor (EGF), insulin-like growth factors (IGFs), transforming growth factor beta (TGF-β), vascular endothelial growth factor (VEGF), and others [ 19–21 ]. These growth factors play important roles in cell proliferation, migration, and differentiation. Our previous study revealed that activated PRP has a potential effect on the proliferation of hASCs and human dermal fibroblasts (hDFs) compared with nonactivated PRP [ 22 ]. Furthermore, we also reported that activated PRP induces hDF proliferation via the activation of ERK1/2 signaling [ 23 ]. Recently, other investigators reported that PDGF also enhances proliferation of hASCs through the JNK pathway [ 24 ]. However, the signaling pathways involved in PRP-stimulated proliferation of hASCs have not been clarified. In the present study, we show that PRP stimulated cell proliferation by ERK1/2, JNK, and Akt activation. We compared this effect with the proliferative effect of PDGF-BB, a major growth factor in PRP. Methods Preparation of activated PRP Activated PRP was obtained using the double-spin method as described previously [ 23 ]. Briefly, after obtaining informed consent from healthy adult volunteers (n = 3), blood was collected in tubes containing an acid-c (...truncated)


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Fangyuan Lai, Natsuko Kakudo, Naoki Morimoto, Shigeru Taketani, Tomoya Hara, Takeshi Ogawa, Kenji Kusumoto. Platelet-rich plasma enhances the proliferation of human adipose stem cells through multiple signaling pathways, Stem Cell Research & Therapy, 2018, pp. 107, Volume 9, Issue 1, DOI: 10.1186/s13287-018-0851-z