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시험관 내 면역 기술에 의한 항체 생산에 관한 연구

  • 전태훈;서동상
    • The Korean Journal of Zoology
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    • v.37 no.1
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    • pp.19-32
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    • 1994
  • 가장 효과적인 H-2b 항원에 대한 시험관내 항체 생산 조건을 찾기 위하여 근교계 생쥐인 C57BL/6BySnj의 비장세포를 UV로 불활성학 시킨 후 항원으로 사용하고, A/wySnJ$\times$Sm/J(ASmJF1, hybrid)의 비장세포를 항원 수용자 계통으로 하여 5-7일동안 배양기에서 항체 생산을 유도하였다. 본 실험에서 T 임파구 대식세포와 임파구 분화 촉진 인자인Concanavalin A Lipopolvsaccharide. Pokeweed mitogen 등을 사용하여 20가지 조건으로 실험을 수행하여, 항체 생성 여부는 보체 의존성 세포 장애 실험과 면역 효소법에 의해 조사하였다 그 결과 모든 조건에서 항체생산이 확인되었으며. 가장 좋은 시험관내 항체 생산 조건으로는 T 임파구와 대식세포를 함께 사용하여 면역시킨 것이 가장 효과적이었다. 이 방법을 이용하여 항체 생산을 유도한 후 5일째 면역된 비장세포를 Sp2/0-Ag 14와 세포 융합시켜 H-2b 마우스의 체포 표면 항원에 대한 단일군항체 생산을 시도하였다. 또한 생체내 면역 방법과 비교하기 위해 6주간 C57BL/6BySnJ의 비장세포를 복강내에 주사하여 같은 조건으로 세포융합을 시도하였다. 그 결과 H-2b 세포의 표면 항원에 대한 항체 생산을 하는 세포군은 시험관내 면역 방법에서 3개 생체내 면역 방법에서 4개부 확인되었다.

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Effect of Di-(2-ethylhexyl)phthalate(DBHP) on Spermatogenesis in Rat Testes (흰쥐 정자형성과정에 미치는 Di-(2-ethylhexyl)phthalate의 영향)

  • 김완종;길영천;이종화;신길상
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.285-292
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    • 1999
  • Di-(2-ethylhexyl) phthalate (DEHP) is a plasticizer known as one of endocrine disruptors. The present study was carried out to investigate the alterations of function and ultrastructure in rat testes after oral intubation of DEHP in dosages of 1g/kg/day, 2 g/kg/day or 3 g/kg/day in 0.5 ml of corn oil for 15 days. DEHP reduced the growth of body and testes, inhibited apermatogenesis and induced structural changes on various cell types of the rat testis. Leydig cells, Sertoli cells and the developing germ cells seemed to be impaired their differentiations in terms of the structural changes of cell organelles. The increase of heterochromatin in amount were common features in all 3 cell types. In addition, the Leydig cells were characterized by the swelling of smooth endoplasmic reticulum and perinuclear space, the increases in number and size of Iysosomes. The Sertoli cells became irregular in nuclear envelope and the number of Iysosomes and vacuoles seemed to be increased. There were some indications of necrosis of the germ cells, such as vacuolized nucleus and segregated nucleolus. And also, DEHP lowered the level of testosterone in experimental rat serum. DEHP suppressed apermatogenesis decreasing developing germ cells and these effects of DEHP on the rat testis were dose dependent. The detrimental effect of DEHP on apermatogenesis and ultrastructure of rat testes seems to be derived from the decreased level of testosterone by Leydig cells, followed by the abnomalities of Sertoli cells and the germ cells.

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Spermatozoa Characteristics of Streptozotocin-induced Diabetic Wistar Rat: Acrosome Reaction and Spermatozoa Concentration (Streptozotocin으로 유발된 당뇨병성 Wistar Rat 정자의 첨체반응 및 수 변화 특성)

  • Cheon, Yong-Pil;Kim, Chung-Hoon;Kang, Byung-Moon;Chang, Yoon-Seok;Nam, Joo-Hyun;Kim, Young-Soo;Gye, Myung-Chan;Kim, Moon-Kyoo;Kim, Kil-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.89-96
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    • 1999
  • Some of the information concerning sexual function in the male diabetes has been focused upon the problems of endocrine or semen parameters. However, the characteristics of acrosome reaction and spermatozoa concentration at the epididymis and vas deferens have scarcely been studied, and the causes of the infertility has not been critically identified. So, we designed to inspect the spermatozoa concentration and the characteristics of acrosome reaction at epididymis and vas deferens of diabetic Wistar rat induced by streptozotocin (STZ, 70 mg/kg, ip). Experimental animal was sacrificed at 3 days and 14 days after the STZ injection. In the diabetes-induced rat, the levels of insulin and glucose had a pattern of inverse proportion. The spermatozoa concentrations in caput and corpus epididymis were significantly decreased in all diabetic condition. In cauda epididymis, however, there was significant decrease in sperm concentration at 14 days onward. In diabetic rat, the spontaneous reaction rate of spermatozoa of cauda and vas deferens were significantly higher than the control group. The ARIC (acrosome reaction to ionophore challenge) value of caudal sperm was 28.7 at control, 22.1 at 3 days, and 8.3 at 14 days. In the present study the spermatozoa concentration was decreased and the spontaneous reaction rate was increased by diabetes. In ARIC-test, it is revealed that the fertility of spermatozoa of 14 days group was lower than control or 3 days group. Diabetes mellitus may be provoke the decreased fertilization rate and subsequent infertility and subsequent infertility.

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