title: Activated platelet supernatant can augment the angiogenic potential of human peripheral blood stem cells mobilized from bone marrow by G-CSF.
abstract: Platelets not only play a role in hemostasis, but they also promote angiogenesis and tissue recovery by releasing various cytokines and making an angiogenic milieu. Here, we examined autologous 'activated platelet supernatant (APS)' as a priming agent for stem cells; thereby enhance their pro-angiogenic potential and efficacy of stem cell-based therapy for ischemic diseases. The mobilized peripheral blood stem cells ((mob)PBSCs) were isolated from healthy volunteers after subcutaneous injection of granulocyte-colony stimulating factor. APS was collected separately from the platelet rich plasma after activation by thrombin. (mob)PBSCs were primed for 6h before analysis. Compared to naive platelet supernatants, APS had a higher level of various cytokines, such as IL8, IL17, PDGF and VEGF. APS-priming for 6h induced (mob)PBSCs to express key angiogenic factors, surface markers (i.e. CD34, CD31, and CXCR4) and integrins (integrins α5, β1 and β2). Also (mob)PBSCs were polarized toward CD14(++)/CD16(+) pro-angiogenic monocytes. The priming effect was reproduced by an in vitro reconstruction of APS. Through this phenotype, APS-priming increased cell-cell adhesion and cell-extracellular matrix adhesion. The culture supernatant of APS-primed (mob)PBSCs contained high levels of IL8, IL10, IL17 and TNFα, and augmented proliferation and capillary network formation of human umbilical vein endothelial cells. In vivo transplantation of APS-primed (mob)PBSCs into athymic mice ischemic hindlimbs and Matrigel plugs elicited vessel differentiation and tissue repair. In safety analysis, platelet activity increased after mixing with (mob)PBSCs regardless of priming, which was normalized by aspirin treatment. Collectively, our data identify that APS-priming can enhance the angiogenic potential of (mob)PBSCs, which can be used as an adjunctive strategy to improve the efficacy of cell therapy for ischemic diseases.
authors.affiliation: Department of Internal Medicine, Cardiovascular Center, Seoul National University Hospital, Republic of Korea; Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea.
authors.forename: Jeehoon
authors.initials: J
authors.lastname: Kang
authors.affiliation: Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Jin
authors.initials: J
authors.lastname: Hur
authors.affiliation: Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Jin-A
authors.initials: JA
authors.lastname: Kang
authors.affiliation: Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Ji-Yeon
authors.initials: JY
authors.lastname: Yun
authors.affiliation: Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Jae-Il
authors.initials: JI
authors.lastname: Choi
authors.affiliation: Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Seung Bum
authors.initials: SB
authors.lastname: Ko
authors.affiliation: Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Choon-Soo
authors.initials: CS
authors.lastname: Lee
authors.affiliation: Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Jaewon
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Internal Medicine, Cardiovascular Center, Seoul National University Hospital, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea.
authors.forename: Jung-Kyu
authors.initials: JK
authors.lastname: Han
authors.affiliation: Department of Laboratory Medicine, Seoul National University Hospital, Republic of Korea.
authors.forename: Hyun Kyung
authors.initials: HK
authors.lastname: Kim
authors.affiliation: Department of Internal Medicine, Cardiovascular Center, Seoul National University Hospital, Republic of Korea; Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Republic of Korea; National Research Laboratory for Stem Cell Niche, Republic of Korea. Electronic address: hyosoo@snu.ac.kr.
authors.forename: Hyo-Soo
authors.initials: HS
authors.lastname: Kim
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology*; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008954:Models, Biological; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D019899:Paracrine Communication / Q000187:drug effects; D010641:Phenotype; D015539:Platelet Activation / Q000187:drug effects*; D012038:Regeneration / Q000187:drug effects; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013927:Thrombosis / Q000473:pathology
chemical_list: D016207:Cytokines; D016179:Granulocyte Colony-Stimulating Factor
keywords: Activated platelet supernatant; Angiogenesis; Cell priming; Mobilized peripheral blood stem cells; Stem cell-based therapy