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Differential regulation of mTORC1 and mTORC2 is critical for 8-Br-cAMP-induced decidualization
백미옥,송해인,한중수,윤미섭 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-
Human endometrium decidualization, a differentiation process involving biochemical and morphological changes, is a prerequisite for embryo implantation and successful pregnancy. Here, we show that the mammalian target of rapamycin (mTOR) is a crucial regulator of 8-bromoadenosine 3’,5’-cyclic monophosphate (8-Br-cAMP)-induced decidualization in human endometrial stromal cells. The level of mSin1 in mTOR complex 2 (mTORC2) and DEPTOR in mTOR complex 1 (mTORC1) decreases during 8-Br-cAMP-induced decidualization, resulting in decreased mTORC2 activity and increased mTORC1 activity. Notably, DEPTOR displacement increases the association between raptor and insulin receptor substrate-1 (IRS-1), facilitating IRS-1 phosphorylation at serine 636/639. Finally, both S473 and T308 phosphorylation of Akt are reduced during decidualization, followed by a decrease in forkhead box O1 (FOXO1) phosphorylation and an increase in the mRNA levels of the decidualization markers prolactin (PRL) and insulin-like growth factor-binding protein-1 (IGFBP-1). Taken together, our findings reveal a critical role for mTOR in decidualization, involving the differential regulation of mTORC1 and mTORC2.
천연재료가 함유된 조직공학용 지지체에 대한 세포독성 분석법의 비교
백미옥 ( Mi Ock Baek ),김순희 ( Soon Hee Kim ),소정원 ( Jung Won So ),노혜원 ( Hye Won Roh ),이나리 ( Na Ri Lee ),김문석 ( Moon Suk Kim ),유규하 ( Gyu Ha Ryu ),조양하 ( Yang Ha Cho ),이승진 ( Seung Jin Lee ),강길선 ( Gilson Khang 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.4
Cytotoxicity measurements on materials are extremely important in tissue engineering field. The objective of this study is to measure and compare the cytotoxicity of scaffold for tissue engineered using 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide(MTT), 2-(methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2-4- disulfophenyl)-H-tetrazolium(WST-8) and Alamar Blue cytotoxicity assay on cytotoxixity of scaffold for tissue engineering. In order to evaluate the sensitivity of rat bone marrow stromal cell(rBMSC) for cytotoxicity assays, a series of two-fold dilution of rBMSC were plated in 96-wells. rBMSC suspension was seeded on poly(lactide-coglycolide) (PLGA) scaffold and then cytotoxicity was measured in vitro by MTT, WST-8 and alamar blue assay. The MTT assay was more sensitive than WST-8, alamarblue assay for rBMSC. rBMSC suspension seeded on various content of demineralized bon particle(DBP) impregnated PLGA scaffold was measured to confirm on affect to natural material by MTT assay. In the MTT assay results, optical density was influenced by content of DBP. Finally, cytotoxicity of DBP/PLGA scaffold was assayed using modified method of ASTM F813-07. We confirmed that cell viability was significantly high in PLGA scaffold including DBP of 20 and 40% compared to other scaffolds. This report may be useful as guideline in tissue engineered scaffolds.
ASTM F2027 조직공학적 의료용제품(TEMPs)을 위한 기질의 특성결정 및 테스트를 위한 표준 지침서
백미옥 ( Mi Ock Baek ),소정원 ( Jung Won So ),김순희 ( Soon Hee Kim ),노혜원 ( Hye Won Roh ),이나리 ( Na Ri Lee ),김문석 ( Moon Suk Kim ),유규하 ( Gyu Ha Ryu2 ),조양하 ( Yang Ha Cho ),이승진 ( Seung Jin Lee ),강길선 ( Gilson Khang 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.4
Tissue engineering medical products are those protocols and products developed for use in the human body as biological substitutes to restore, maintain, or improve tissue function. The purpose of this standard is to locate relevant existing guideline and test methods and to provide guidance for interim use of materials for which a standard does not exist. The standard may be use as guideline in tissue engineered research.
탈미네랄화된 골분용액을 함침시킨 PLGA 지지체의 제조 및 특성
백미옥 ( Mi Ock Baek ),김순희 ( Soon Hee Kim ),소정원 ( Jung Won So ),임지예 ( Ji Ye Lim ),최진희 ( Jin Hee Choi ),이종문 ( John M. Rhee ),이해방 ( Hai Bang Lee ),강길선 ( Gilson Khang ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.2
We developed the synthetic/natural hybrid scaffolds with poly (lactide-co-glycolide)(PLGA) and solution using DBP(DB solution) for tissue engineering. Composite scaffolds of PLGA scaffold penetrated DB solution (PLGA-pen-DBP) were manufactured by simple method that PLGA scaffolds manufactured by solvent casting/salt leaching method soaked in DB solution. We evaluated pore structure, wettability, water uptake ability and cell viability of two types of scaffolds. Through SEM and FT-IR, it was confirmed that DB solution was put into the pores of PLGA scaffold. Also, the effects of the impregnation of DB solution on PLGA scaffolds were characterized through water uptake ability and cell viability. It seemed that DBP provided proper pore to PLGA scaffold and improved water absorption. A novel scaffold, which PLGA scaffold penetrated with DB solution, might be suitable to apply as tissue engineered scaffold.
PLGA 지지체에 함유된 DBP 함량이 줄기세포의 골분화에 끼치는 영향에 대한 in vitro 테스트
백미옥 ( Mi Ock Baek ),김순희 ( Soon Hee Kim ),소정원 ( Jung Won So ),임지예 ( Ji Ye Lim ),노혜원 ( Hye Won Roh ),이나리 ( Na Ri Lee ),유규하 ( Gyu Ha Ryu ),조양하 ( Yang Ha Cho ),이승진 ( Seung Jin Lee ),강길선 ( Gil Son Khang ) 한국조직공학과 재생의학회 2008 조직공학과 재생의학 Vol.5 No.1
Demineralized bone particles(DBP) exhibit osteoinductive properties and are very useful for bone formation and other orthopeadic applications. In this study, we investigated the osteogenic capabilities of rat bone marrow stromal cells(rBMSCs) in PLGA scaffold impregnated with various DBP contents. DBP-loaded PLGA scaffold (10, 20, 40, 80 % w/w) were prepared by solvent casting/salt leaching method. rBMSCs were seeded in PLGA and DBP/PLGA scaffolds, and then cellular viability and proliferation were assayed by water-soluble tetrazolium salt(WST-8) assay. Osteoblast differentiation of cells were analyzed by alkaline phosphatase(ALP) activity. Finally, reverse transcript polymerase chain reaction(RT-PCR) was assessed to measure mRNA expression of the genes (ALP, osteocalcine and type I collagen) of osteogenic marker in rBMSCs depending on various content of DBP. These results could demonstrated that presence of DBP in PLGA might play important roles in osteoblast differentiation of rBMSCs.