• Title/Summary/Keyword: P4-BSA-FITC

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Synthesis of Methoxy Poly(ethylene glycol)-b-poly($\varepsilon$-caprolactone) Diblock Copolymers and Release Behavior of Albumin for Implantable Protein Carriers (이식형 단백질 전달체로서 메톡시 폴리(에틸렌 글리콜)/폴리카프로락톤 블록 공중합체의 합성 및 알부민의 방출 거동)

  • 서광수;전세강;김문석;조선행;이해방;강길선
    • Polymer(Korea)
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    • v.28 no.3
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    • pp.232-238
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    • 2004
  • MPEG-PCL diblock copolymers consisting of methoxy poly(ethylene glycol) (MPEG) and $\varepsilon$-caprolactone (CL) as drug carriers were synthesized by ring-opening polymerization MPEG-PCL diblock copolymers were characterized by X-ray diffraction and differential scanning calorimetry. After freeze milling of block copolymers and albumin bovine-fluorescein isothiocyanate (FITC-BSA) as model protein, the wafers loaded FITC-BSA were fabricated by direct compression method. The release profiles of FITC-BSA were examined using pH 7.4 PBS for 14 days at 37$^{\circ}C$. The release amount was determined by fluorescence intensity by using the fluorescence spectrophotometer. The morphological change of wafers was observed by digital camera and scanning electron microscope. The release rate and initial burst of BSA increased with increasing PEG molecular weights and decreasing PCL molecular weights in the segments of MPEG -PCL diblock copolymers.

Preparation and Release Behavior of Methoxy poly(ethylene glycol)- poly(L-lactide-co-glycolide) Wafer Containing Albumin (알부민을 함유한 메톡시 폴리(에틸렌 글리콜)- 폴리(L-락타이드-co-글리콜라이드) 웨이퍼의 제조 및 방출거동)

  • 서광수;김문석;김경자;조선행;이해방;강길선
    • Polymer(Korea)
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    • v.28 no.4
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    • pp.328-334
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    • 2004
  • A series of methoxy poly(ethylene glycol) (MPEG)-poly(L-lactide-co-glycolide) (PLGA) diblock copolymers were synthesized by ring-opening polymerization of L-lactide and glycolide with carbitol (134 g/mole) or different molecular weights of MPEG (550, 2000, and 5000 g/mole) as an initiator in presence of Sn(Oct)$_2$. The properties of diblock copolymers were characterized by using $^1$H-NMR, GPC, and XRD. After uniform mixing of block copolymers and 1% albumin bovine-fluorescein isothiocyanate(FITC-BSA) with a freeze miller, the wafers loaded FITC-BSA were fabricated by using a mold with a dimensions of 3 mm${\times}$1mm diameter. The release profiles of FITC-BSA and the pH changes of wafer were examined using pH 7.4 PBS for 30 days at 37$^{\circ}C$. The release profiles of albumin showed fast initial burst as the molecular weights of MPEG increased. As a result of this study, the release behavior of BSA was controlled with introducing MPEG in the block copolymers.

Preparation and Release Behavior of Albumin-Loaded PLGA Scaffold by Ice Particle Leaching Method (얼음입자추출법을 이용한 알부민 함유 PLGA 담체의 제조 및 방출 거동)

  • Hong Keum Duck;Seo Kwang Su;Kim Soon Hee;Kim Sun Kyung;Khang Gilson;Shin Hyung Sik;Kim Moon Suk;Lee Hai Bang
    • Polymer(Korea)
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    • v.29 no.3
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    • pp.282-287
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    • 2005
  • A novel ice particle leaching method for fabrication of porous and biodegradable PLGA scaffold has been proposed for the application to tissue engineering. After uniform mixing of poly(L-lactide-co-glycolide) (PLGA) and bovine serum albumin-fluorescein isothiocyanate (FITC-BSA), the FITC-BSA loaded scaffold was fabricated by adding various ratio of ice particle. The release profiles of FITC-BSA were examined using pH 7.4 PBS for 28 days at $37^{circ}$. The release amount was determined by fluorescence intensity by using the fluorescence spectrophotometer and the morphological change of the scaffolds was observed by scanning electron microscope. The release initial burst of BSA containing scaffolds was lower than that of simple dipping scaffolds resulting in constant release aspect. Although the BSA concentration increased. the initial burst was not increased. As a result of this study, it can be suggested that ice particle leaching method for the tissue engineered scaffold miff be very useful and it is possible to impregnate with water soluble factors like cytokine. We suggest that ice particle leaching method may be useful to tissue engineered organ regeneration.

Expression of Progesterone Receptor Membrane Component 1 and 2 in the Mouse Gonads and Embryos (생쥐 생식소 및 배아의 프로게스테론 수용체 막성분 1과 2의 발현에 관한 연구)

  • Kim, Kyeoung-Hwa;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.11 no.1
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    • pp.21-29
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    • 2007
  • Previously, we found progesterone receptor membrane component 2 (pgrmc2) was highly expressed in germinal vesicle (GV) stage oocytes. The present study was conducted to characterize the expression of pgrmc2, as well as pgrmc1, in the mouse gonads and embryos according to their developmental stages. We found that these membrane components were expressed in ovaries, testes, and embryos at various developmental stages in addition to oocytes. Progesterone-3-O-carboxymethyl oxime-BSA-fluorescein isothiocyanate (P4-BSA-FITC) was applied to visualize the presence of the progesterone receptor on mouse oocyte membrane, and we confirmed that immobilized progesterone is localized at surface of the oocyte. This is, at our knowledge, the first report regarding the expression of membrane component of progesterone receptor in the mouse oocytes, embryos, and gonads. The function and signal transduction pathway of progesterone receptor membrane components in oocytes requires further studies.

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Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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