• 제목/요약/키워드: stromal cells

검색결과 379건 처리시간 0.105초

자궁내막 세포의 분리 및 배양 그리고 불멸화된 세포주의 확립 (Endometrial Cell Culture: Isolation, Characterization, and Immortalization)

  • 홍인선;김석현;궁미경;전진현;이영순;강경선
    • Clinical and Experimental Reproductive Medicine
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    • 제30권4호
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    • pp.317-324
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    • 2003
  • 목 적: 본 실험의 목적은 자궁내막세포를 분리 및 배양법 확립과 함께 불멸화 시키는 것이다. 방 법: 자궁내막에서 상피세포(epithelial cells)와 기질세포(stromal cells)의 분리는 Satyawaroop 등(1979)의 방법에 기초를 두었다. 자궁내막에서 상피세포와 기질세포의 순수 분리도를 확인하고, 불멸화된 기질세포에서 SV40 large T antigen을 확인하기 위하여 면역형광 염색(immunocytochemistry)과 Western blot 기법을 이용하였다. 정상 기질세포의 경우 subconfluence (60%) 상태에서 transfection을 진행하였다. 순수 분리된 plasmid DNA와 Qiagen 사의 superfect를 이용하여 transfection을 실시하였다. 결 과: 본 연구에서 우리는 두 가지 형태의 자궁내막 세포의 분리 및 배양에 성공하였다. 상피세포는 다면체의 형태를 띠며, 선(grandular)조직의 조각으로부터 나선형으로 자란다.기질 세포는 길쭉한 형태를 띠며, 상피세포에 비해 오래 살고, 빠르게 증식하여 나란한 형태로 배열된 세포 다발(cell bundle)을 형성한다. 이렇게 분리된 세포들은 95%의 균질성을 보였으며, 면역형광염색과 western blot을 통해 확인 하였다. 한편 SV40(Simian Virus 40) large T 항원을 암호화 하고 있는 염기 서열을 포함한 플라스미드 벡터로 안정적인 트랜스펙션을 시킴으로써 불멸화 된 자궁내막의 기질 세포주를 확립하였다. 불멸화 된 세포는 그 세포가 유래한 정상의 세포와 동일한 표현형을 가지고 있었다. 결 론: 본 연구에서, 우리는 자궁내막에서 상피세포(epithelial cells)과 기질세포(stromal cells)를 분리하여 배양법을 확립하였다. 동시에 SV40 large T antigen을 이용하여 불멸화된 세포주를 확립하였다. 이렇게 확립된 세포주는 자궁의 생리작용 연구 및 자궁내막증(Endometriosis)과 자궁암(Endometrial cancer) 등과 같은 여러 자궁관련 질병 연구에 많은 도움이 될 것으로 사료된다.

황백이 만성 비세균성 전립선염 모델에서 혈액학적 및 세포조직학적 변화에 미치는 영향 (The Effects of Phellodendri Cortex Treatment on Hematological and Cyto-pathological Alterations in Non-Bacterial Prostatitis Rat Model)

  • 김순일;안영민;안세영;두호경;이병철
    • 대한한의학회지
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    • 제27권3호
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    • pp.51-62
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    • 2006
  • Objective: Although chronic non-bacterial prostatitis is a common disease, it is very difficult to treat effectively. Lygodium japonicum has traditionally been used in treatment of urinary tract inflammation and voiding disturbance. In this study, we investigated the therapeutic effects and action mechanism of Lygodium japonicum in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods: Five-month-old rats were treated with $17\beta-estradiol$ after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histopathological profiles. Lygodium japonicum and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histopathological parameters including the epithelial score and epithelio-stromal ratio for glandular damage, proliferating cell nuclear antigen (PCNA) labeling index for cyto-proliferation and a TUNEL (deoxyuridine triphosphate biotin nick end-labeling) assay for cell apoptosis. Results: While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Lygodium japonicum showed a lesser range of tissue damage. Epithelial score was improved in Lygodium japonicum than that of the control (P<0.05). The epithelio-stromal ratio was lower in Lygodium japonicum when compared to that of the control (P<0.05). Although there was no difference in PCNA and TUNEL positive cells of the glandular epithelia, we found an decreased number of PCNA positive cell and concurrent increase of TUNEL positive cells in the stroma of Lygodium japonicum treated rats (P<0.01). Conclusions: These findings suggest that Lygodium japonicum may protect the glandular epithelial cells and also inhibit stromal proliferation in association with suppression of cyto-proliferation and stimulation of apoptosis. We concluded that Lygodium japonicum may be a useful remedy agent for treating the chronic non-bacterial prostatitis.

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흰쥐 가슴샘 재생과정 동안 대식세포에서 SDF-1의 발현증가 (Expression of Stromal Derived Factor-1 is Upregulated In Macrophages during Thymic Regeneration in Adult Rat)

  • 박현주;김종갑;윤식;배문경;배수경
    • 생명과학회지
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    • 제19권8호
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    • pp.1067-1072
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    • 2009
  • 성체흰쥐의 경우 항암제인 싸이클로포스파마이드(CY)처리로 퇴축된 가슴샘은 2주 후에 정상조직으로 재생된다. 가슴샘 발생과정에서 이미 알려진 SDF-1 신호전달의 중요성과는 달리 성체의 가슴샘 재생과정에서 그 역할에 관해서는 알려진 바 전혀 없다. 본 연구의 목적은 발생중인 가슴샘에서 발현이 증가된다고 이미 알려져 있는 SDF-1이 성체의 가슴샘재생과정에서 어떤 발현 양상을 보이는지를 조사하는 것이다. 본 연구에서는 사이클로포스파마이(cyclophosphamide: CY)를 투여하여 가슴샘의 급성 퇴축을 유발시킨 후, 가슴샘 재생과정동안 SDF-1의 발현 특성을 면역조직화학, 이중면역염색 형광법, 역전사 중합효소 연쇄반응법을 이용하여 조사하였다. 그 결과, SDF-1은 가슴샘의 급성 퇴축 이후 7일째 되는 시기에 mRNA와 단백질의 양이 급격히 증가하였으며, 이중면역염색 형광법을 통해 큰포식 세포와 위치적 분포가 일치함을 확인하였다. 따라서, 본 연구 결과들을 통해 SDF-1은 가슴샘의 급성 퇴축 초기 과정에서 나타나는 손상된 세포를 처리하는 큰포식 세포의 기능 조절에 관여할 것으로 생각된다.

해금사가 만성 비세균성 전립선엽 모델에서 혈액학적 및 세포조직학적 변화에 미치는 영향 (The Effects of Lygodium japonicum Treatment on Hematological and Cyto-pathological Alterations in Non-Bacterial Prostatitis Rat Model)

  • 이병철;김상우;안영민;두호경;안세영
    • 대한한방내과학회지
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    • 제27권3호
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    • pp.664-676
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    • 2006
  • Objective : Although chronic non-bacterial prostatitis is a common disease, it is very difficult to treat effectively. Lygodium japonicum has been traditionally used in treatment of urinary tract inflammation and voiding disturbance. In this study, we investigated the therapeutic effects and action mechanism of Lygodium japonicum in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods : Five-month-old rats were treated with 17$\beta$-estradiol after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histopathological profiles. Lygodium japonicum and testosterone were administered as an experimental specimen and a positive control. respectively. The prostates were evaluated by histopathological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. PCNA labeling index for cyto-proliferation and a TUNEL(deoxyuridine triphosphate biotin nick end-labeling) assay for cell apoptosis. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Lygodium japonicum showed a diminished range of tissue damage. Epithelial score was improved in the Lygodium japonicum group over that of the control (P<0.05). The epithelio-stromal ratio was lower in the Lygodiutn japonicum group when compared to that of the control (P<0.05). Although there was no difference in PCNA and TUNEL positive cells of the glandular epithelia. we found an decreased number of PCNA positive cell and concurrent increase of TUNEL positive cells in the stroma of Lygodium japonicum treated rats (P<0.01). Conclusions : These findings suggest that Lygodium japonicum may protect the glandular epithelial cells and also inhibit stromal proliferation in association with suppression of cyto-proliferation and stimulation of apoptosis. We concluded that Lygodium japonicum could be a useful remedy agents for treating chronic non-bacterial prostatitis.

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커큐민과 지방 기질세포가 흰 쥐의 창상치유에 미치는 영향 (The Effect of Curcumin and Human Adipose-derived Stromal Cells on Wound Healing of Lewis Rats)

  • 백혜원;황동연;유결;변준희
    • Archives of Plastic Surgery
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    • 제37권2호
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    • pp.110-114
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    • 2010
  • Purpose: On this study, we investigated the effects of curcumin and adipose-derived stromal cells (ADSCs) in wound healing process, especially in the aspect of synergic effects when they were administrated simultaneously. Methods: Curcumin (40 mg/kg) and/or $1.0{\times}10^6$ ADSCs were applied to an $1.5{\times}1.5\;cm$-sized full thickness wound on the backs of male Lewis rats (n=5 in each group). In control group (n=5), saline was administrated instead of curcumin and ASCs. The wound size was followed by computer planimetry in 5, 7, and 14 days, and wounds were harvested for histological analysis in 7 and 14 days. Results: The dimensions of wounds of curcumin, ADSCs, and curcumin-ADSCs group significantly decreased in 5, 7, 14 days compared with those of control group (p<0.05), but there were no significant differences among three groups. The wound sizes were lowest in curcumin-ADSCs group compared with the other groups, but the differences were insignificant (p>0.05). There were infiltration of more epithelization and more precisely organization of extracellular matrix in curcumin, ADSCs, and curcumin-ADSCs group compared with those of control group. Conclustion: The results suggest that curcumin and ADSCs have beneficial effects in the acceleration of wound healing. Although the simultaneous application of curcmin and ADSCs also has beneficial effects on wound healing, there are no significant synergic effects.

인동소염방(忍冬消炎方)이 만성 비세균성 전립선염 Rat 모델에서 혈액 및 세포조직의 변화에 미치는 영향 (The Effects of IDS(Indongsoyeom-bang) Treatment on the Hematological and Cytopathological Alterations in Non-Bacterial Prostatitis Rat Model)

  • 이종빈;황석연;조충식
    • 혜화의학회지
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    • 제20권1호
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    • pp.91-104
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    • 2011
  • Background : Although chronic non-bacterial prostatitis is increasing, it is hard to treat effectively. In western medicine, antimicrobials drug, ${\alpha}$-adreno-ceptor antagonists, anti-inflammatory drugs, tricyclic antidepressants and anticholinergic agents are used commonly, but chronic prostatitis/chronic pelvic pain syndromes is confusing and frustrating for urologist. IDS(Indongsoyeom-bang) is used in treatment of chronic prostatitis/chronic pelvic pain syndromes. And it is reported that GLS(Gleditsiae spina) and TOF(Toosendan fructus) components of IDS have significant effect on protection of the glandular epithelial cells. Objective : In this study was conducted to investigate the therapeutic effects and action machanism of IDS in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods : We observed six experimental objects of normal group, control group, testosterone group, and IDS 50 mg/kg, 200mg/kg, 400mg/kg group. Rats were treated with 17 ${\beta}$-estradiol after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histophatological profiles. IDS and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. Also, the prostates were observed by Hematological alterations of WBC, RBC, hemoglobin and platelet. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with IDS-50 showed a diminished range of the tissue damage. Epithelial score was improved in IDS than that of the control. The epithelio-stromal ratio was lower in IDS when compared to that of the control. Also, the examination of bloods were not observed hematological change. Conclusion : These finding suggests that IDS may protects the glandular epithelial cells. We concluded that IDS could be a useful remedy agent for treating chronic non-bacterial prostatitis.

Enhanced bone morphogenic protein adenoviral gene delivery to bone marrow stromal cells using magnetic nanoparticle

  • Lee, Jung-Tae;Jung, Jae-Whan;Choi, Jae-Yong;Kwon, Tae-Geon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제39권3호
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    • pp.112-119
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    • 2013
  • Objectives: This study investigated the question of whether adenoviral magnetofection can be a suitable method for increasing the efficacy of gene delivery into bone marrow stromal cell (BMSC) and for generation of a high level of bone morphogenic protein (BMP) secretion at a minimized viral titer. Materials and Methods: Primary BMSCs were isolated from C57BL6 mice and transduced with adenoviral vectors encoding ${\beta}$ galactosidase or BMP2 and BMP7. The level of BMP secretion, activity of osteoblast differentiation, and cell viability of magnetofection were measured and compared with those of the control group. Results: The expression level of ${\beta}$ galactosidase showed that the cell transduction efficiency of AdLacZ increased according to the increased amount of magnetic nanoparticles. No change in cell viability was observed after magnetofection with 2 ${\mu}L$ of magnetic nanoparticle. Secretion of BMP2 or BMP7 was accelerated after transduction of AdBMP2 and 7 with magnetofection. AdBMP2 adenoviral magnetofection resulted in up to 7.2-fold higher secretion of BMP2, compared with conventional AdBMP2-transduced BMSCs. Magnetofection also induced a dramatic increase in secretion of BMP7 by up to 10-fold compared to the control. Use of only 1 multiplicity of infection (moi) of magnetofection with adenoviral transduction of AdBMP2 or AdBMP7 resulted in significantly higher transgene expression compared to 20 moi of conventional adenoviral transduction. Conclusion: Magnetic particle-mediated gene transudation is a highly efficient method of gene delivery to BMSCs. Magnetofection can lower the amount of viral particles while improving the efficacy of gene delivery.