• Title/Summary/Keyword: Matrigel

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Development of Mouse Preimplantation Embryos in Solubilized Matrigel Media (용해된 Matrigel 첨가 배지에서 착상전 생쥐 배아의 발생)

  • Chung, Byung-Mok;Choo, Hyung-Sik;Kang, Byung-Moon;Gye, Myung-Chan
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.381-385
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    • 2000
  • Objective: To verify the effect of two forms (growth factor and growthfactor-reduced) of solubilized Matrigel on the development in mouse preimplantation embryos. Methods: Late 2-cell stage eggs were cultured through the blastocyst stage in the presence of GF- or GFR-Matrigel (0.5%, v/v). Morphological development, cell number and % apoptotic nuclei of blastocyst were measured by Roecst staining and TUNEL of nuclei. Results: Morphological development, number of cells per embryo was significantly increased in the presence of GF- or GFR-Matrigel. Culture of the embryos in the GF-Matrigel gave the best result. Conclusion: Low concentration of solubilized Matrigel improved development of mouse embryos regardless of growth factor content of the Matrigel. Growth factors and extracellular matrix protein included in the Matrigel synergistically potentiated the development of mouse embryos.

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Regulation of Mitogen Activated Protein Kinase Activity by Solubilized Matrigel in the Preimplantation Mouse Embryos (생쥐 착상 전 배아에서 용해된 Matrigel에 의한 Mitogen Activated Protein Kinase 활성의 조절)

  • 강병문;정병목;계명찬
    • Development and Reproduction
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    • v.6 no.1
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    • pp.1-6
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    • 2002
  • To elucidate the mechanism underlying the embryotropic effect of extracellular matrix(ECM) on the preimplantation development of mammalian embryos, the involvement of mitogen-activated protein kinase(MAPK) downstream the integrin signaling was examined in mouse blastocysts. Blastocysts were cultured in the presence of growth factor-reduced(GFR) Matrigel(0.5%, v/v). MAPK activity was measured by in vitro phosphorylation of myelin basic protein by the Erk1/2 antibody immunoprecipitates of embryonic extract following the Matrigei treatment. MAPK activity of the early blastocysts rapidly increased within 10 min fo1lowing the Matrigel treatment. When the embryos were cultured for 12 h in the presence of Matrigel, the MAPK activity was significantly higher than that ot the control embryos. PD098059, a MAPK kinase(MEK) inhibitor, attenuated the effect of Matrigel on the change in MAPK activity. Taken together, it suggested that the embryotropic effect of ECM proteins might be mediated by the activation of MAPK cascade.

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Development of In vitro Fertilized Bovine Embryos in Medium Supplemented with Matrigel (Matrigel이 첨가된 배양액에서 소 체외수정란의 발달)

  • 김동훈;김세웅;이민정;황인선;배성훈;양병철;임기순;성환후;양보석
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.805-812
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    • 2006
  • The purpose of this study was to investigate the effect of Matrigel on the development of bovine embryos after in vitro fertilization. Bovine embryos were cultured in Ⅰ) SOF+ 0.8% BSA(SOF-B), Ⅱ) SOF+ 0.8% BSA plus 0.8% Matrigel(SOF-M) and III) SOF+0.8% BSA and 10% FBS(SOF-BF). The addition of Matrigel appeared not to increase the proportion of blastocysts (SOF-B, 26.6%; SOF-M, 28.2%; SOF-BF, 26.2%). However, the proportion of hatched blastocysts were significantly increased(P<0.05) by Matrigel(SOF-B, 23.7%; SOF-M, 48.7%; SOF-BF, 18.5%). The means of cell number blastocyst was not significantly different among the treatment groups(SOF-B, 172.7±35.5; SOF-M, 175.1±37.4; SOF-BF, 172.8±38.1). The proportion of apoptotic cells in blastocyst was also found to be not significant among the treatment groups(SOF-B, 3.6±3.2%; SOF-M, 4.3±2.6%; SOF-BF, 4.9±4.3%). In this experiment, Matrigel appeared to support embryonic hatching of bovine embryos. Results suggest that Matrigel, as extracellular matrix components, may be another avenue for formulating more physiological culture system in serum-free culture.

Regulation of Preimplantation Development of Mouse Embryos by Solubilized Matrigel (용해된 Matrigel에 의한 생쥐 초기배아 발생의 조절)

  • 계명찬;정병목
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2001.02a
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    • pp.68-70
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    • 2001
  • 착상전 초기배아에서 용해된 Matrigedl에 의한 배아의 형태발생, 세포증식, apoptosis 및 UPK활성의 변화를 조사하였다. Matrigel (0.5%)을 첨가한 배양액에서 체외배양된 2-세포기 배아의 형태발생 및 포배당 세포수가 증가되었으며 (GF>GFR>control) 포배의 TUNEL 양성 할구 및 배아내 caspase-3의 활성이 감소되었다. (GF

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Preimplantation Development and Apoptosis of Mouse Embryos in the Medium Containing Extracellular Matrix (Extracellular Matrix 배지에서 생쥐 배아의 발생 및 아폽토시스)

  • Kang, Byung-Moon;Sohn, In-Pyo;Chung, Byung-Mok;Choi, Kyoo-Wan;Gye, Myung-Chan
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.253-259
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    • 2000
  • Objective: To verify the effect of Matrigel, a ECM complex from Engelbreth-Holm-Swarm (EHS) mouse sarcoma on the preimplantation development and apoptosis of mouse fertilized eggs. Method: Late pronucleus stage eggs were cultured through the blastocyst stage in the presence of Matrigel (0.5%, v/v). Characteristics of apoptosis and cell number assesed by Hoecst staining and TUNEL labeling at the blastocyst stage, respectively. Results: Morphological development, number of cells per embryo was significantly increased but rate and number of TUNEL positive nuclei of the embryo were decreased in the presence of Matrigel. Conclusion: This result suggested that at low concentration of Matrigel improves both viability and morphological development in the preimplantation mouse embryos.

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Effects of Kimchi Extract on the Development of Multicellular Structures from Rat Mammary Organoids Cultured in Matrigel

  • Kim, Nam-Deuk;Hur, Young-Mi;Rhee, Sook-Hee;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.168-173
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    • 1996
  • The effect of methanol soluble fraction(MSF) of kimchi on the proliferating and differentiating activity of normal rat mammary epithelial cells or organoids in culture were studied. Reconstituted basement membrane, Matrigel, supported the growth and development several different multicellular structures from mammary organoids. The five type colonies of multicellular structures, stellate, ductal, webbed, squamous, and lobulo-ductal colonies, were observed in Matrigel culture. In methanol extract groups, webbed colonies were more and squamous colonies were less than control group. and the lobulo-ductal colonies which is known that it formed in well differentiated mammary epoithelial cells were developed more in MSF treated group than control group. These results showed that methanol extract of kimchi affected on the proliferation and differentiation of normal rat mammary epithelial cells cultured in serum free medium condition.

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Effect of Matrigel for Bone Graft using Hydroxyapatite/Poly $\varepsilon$-caprolactone Scaffold in a Rat Calvarial Defect Model (랫드의 두개골결손부 모델에서 HA/PCL 지지체를 사용한 골이식 시 Matrigel의 효과)

  • Kim, Se-Eun; Shim, Kyung-Mi;Kim, Seung-Eon;Choi, Seok-Hwa;Bae, Chun-Sik;Han, Ho-Jae;Kang, Seong-Soo
    • Journal of Veterinary Clinics
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    • v.27 no.4
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    • pp.325-329
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    • 2010
  • The osteogenic potential of hydroxyapatite/poly $\varepsilon$-caprolactone composite (HA/PCL) scaffolds with matrigel was evaluated in a rat calvarial defect model. Calvarial defect formation was surgically created in Sprague Dawley rats (n = 18). HA/PCL scaffold was grafted with matrigel (M-HA/PCL group, n = 6) or without matrigel (HA/PCL group, n = 6). A critical defect group (CD group, n = 6) did not received a graft. Four weeks after surgery, bone formation was evaluated with radiography, micro computed tomography (micro CT) scanning, and histologically. No bone tissue formation was radiographically evident in the CD group. Bone tissue was radiographically evident in the HA/PCL and M-HA/PCL groups, however, there was more bone-similar opacity in the M-HA/PCL group. Micro CT analysis revealed that the bone volume of the M-HA/PCL group was higher than the HA/PCL group, however, no significant difference was found between the HA/PCL and M-HA/PCL groups. Bone mineral density in the M-HA/ PCL group was significantly higher than in the HA/PCL group (p < 0.05). Histologically, new bone was formed only from existing bone in the CD group, showing concavity without bone formation in the defect. In the HA/PCL group, new bone formation was only derived from existing bone, while in the M-HA/PCL group the largest bone formation was observed, with new bone tissue forming at the periphery of existing bone and around the HA/PCL scaffold with matrigel. The results indicate that the combination of HA/PCL scaffold with matrigel may be an effective means of enhancing bone formation in critical-sized bone defects.

Development of the Three-Dimensional Perfusion Culture Technology for the Salivary Ductal Cells (타액선 도관세포의 관류 배양 기술 개발)

  • Kim, Ji Won;Kim, Jeong Mi;Choi, Jeong-Seok
    • International journal of thyroidology
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    • v.11 no.2
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    • pp.160-166
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    • 2018
  • Background and objectives: Salivary hypofunction is one of the common side effects after radioiodine therapy, and its pathophysiology is salivary ductal stenosis resulting from ductal cell injury. This study aimed to develop the functional culture environment of human parotid gland ductal cells in in vitro three-dimensional perfusion culture system. Materials and Methods: We compared plastic dish culture method and three-dimensional culture system containing Matrigel and nanofiber. Morphogenesis of reconstituted salivary structures was assessed by histomorphometry. Functional characteristics were assessed by immunohistochemistry and reverse transcription polymerase chain reaction (aquaporin 5, CK7, CK18, connexin 43, and p21). In addition, we designed the media perfusion culture system and identified higher rate of cell proliferation and expression of connexin 43 in perfusion system comparing to dish. Results: Human parotid ductal cells were well proliferated with the ductal cell characters under environment with Matrigel. In the presence of Matrigel, aquaporin 5, CK18 and connexin 43 were more expressed than 2D dish and 3D nanofiber setting. In the media perfusion culture system, ductal cells in 3D culture media showed higher cells count and connexin 43 expression compared to 2D dish. Conclusion: This in vitro ductal cell perfusion culture system using Matrigel could be used to study for radioiodine induced sialadenitis model in vivo.

Establishment and characterization of porcine mammary gland epithelial cell line using three dimensional culture system (3차원 배양 시스템을 이용한 돼지 유선 상피 세포 주 특성과 설정)

  • Chung, Hak-Jae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.10
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    • pp.551-558
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    • 2017
  • To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

Effects of Astragalus Membranaceus on Angiogenesis (황기가 혈관 형성에 미치는 영향)

  • Seo, Dong-Min;Choi, Do-Young;Lee, Jae-Dong
    • Journal of Acupuncture Research
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    • v.24 no.2
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    • pp.113-123
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    • 2007
  • 목적 : 황기가 혈관 신생 작용이 있는지에 관하여 관찰한다. 황기는 상처의 치유나 허혈성 질환에 효과를 나타내는 것으로 알려져있다. 이러한 효과가 황기의 혈관 신생작용과의 관련성을 이해하며 향후 임상에 쓰일 수 있는 황기 약침액 개발을 위한 기초 자료를 목표로 한다. 방법 : 황기의 혈관 신생 작용의 관찰을 위하여 human umbilical vein endothelial cells(HUVECs)와 Matrigel angiogenesis model을 이용하여 연구하였다. 결과 : 황기는 용량에 따라서 HUVECs의 증식을 나타내었다. 또한 혈관 내피 세포의 이동과 관형 형성을 보였다. 혈관 신생 물질인 basic fibroblast growth factor(bFGF)가 황기에 의해 증가하였다. Matrigel angiogenesis model에서 황기는 조직학적으로 혈관 형성을 촉진하였으며,헤모글로빈의 증가를 나타내었다.

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