S-Space Graduate School of Convergence Science and Technology (융합과학기술대학원) Dept. of Molecular and Biopharmaceutical Sciences (분자의학 및 바이오제약학과) Theses (Master's Degree_분자의학 및 바이오제약학과)
Activated platelet supernatant can augment the angiogenic potential of human peripheral blood stem cells mobilized from bone marrow by G-CSF
활성화 혈소판 상등액을 이용한 G-CSF 동원 말초혈액 자가줄기세포의 혈관 형성능 향상에 관한 연구
- 융합과학기술대학원 분자의학 및 바이오제약학과
- Issue Date
- 서울대학교 대학원
- Activated platelet supernatant; G-CSF; Peripheral blood stem cells; Cell priming; Angiogenesis; Stem cell-based therapy
- 학위논문 (석사)-- 서울대학교 융합과학기술대학원 : 분자의학 및 바이오제약학과, 2015. 2. 김효수.
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 (mobPBSCs) 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. mobPBSCs were primed for 6 hours 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 6 hours induced mobPBSCs to express key angiogenic factors, surface markers (i.e. CD34, CD31, and CXCR4) and integrins (integrin α5, β1 and β2). Also mobPBSC 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 mobPBSCs 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 mobPBSCs into athymic mice ischemic hindlimbs and Matrigel plugs elicited vessel differentiation and tissue repair. In safety analysis, platelet activity increased after mixing with mobPBSCs regardless of priming, which was normalized by aspirin treatment.
Collectively, our data identify that APS-priming can enhance the angiogenic potential of mobPBSCs, which can be used as an adjunctive strategy to improve the efficacy of cell therapy for ischemic diseases.