Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Primary Culture and Characteristics of Blood-Brain Barrier Endothelial Cells from Rat Brain Microvessel

쥐의 뇌 미세혈관 내피세포를 이용한 뇌혈관장벽 내피세포의 배양과 특성

  • Lee, Hee-Sang (Department of Anatomy, College of Medicine, Chung-Ang University) ;
  • Kim, Seok-Jung (Department of Anatomy, College of Medicine, Chung-Ang University) ;
  • Kim, Dae-Jin (Department of Anatomy, College of Medicine, Chung-Ang University) ;
  • Chung, Yoon-Hee (Department of Anatomy, College of Medicine, Chung-Ang University) ;
  • Kim, Sung-Su (Department of Anatomy, College of Medicine, Chung-Ang University) ;
  • Lee, Won-Bok (Department of Anatomy, College of Medicine, Chung-Ang University) ;
  • Kim, Kyung-Yong (Department of Anatomy, College of Medicine, Chung-Ang University)
  • 이희상 (중앙대학교 의과대학 해부학교실) ;
  • 김석중 (중앙대학교 의과대학 해부학교실) ;
  • 김대진 (중앙대학교 의과대학 해부학교실) ;
  • 정윤희 (중앙대학교 의과대학 해부학교실) ;
  • 김성수 (중앙대학교 의과대학 해부학교실) ;
  • 이원복 (중앙대학교 의과대학 해부학교실) ;
  • 김경용 (중앙대학교 의과대학 해부학교실)
  • Published : 2005.12.01
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Abstract

The characteristics of primary cultured rat brain microvessel endothelial cells (RBMECs) were studied using microscopy, immunohistochemistry and measuring of transendothelial electrical resistance (TER). The RBMECs formed a monolayer by $5{\sim}6$ days after plating and showed characteristics of whirling appearance. The TER increased until day 5 and decreased then. There was few immunoreaction with anti-GFAP, anti-GalC, anti-neurofilament 160/200 kD antibodies. So the contamination of astrocyte, oligodendrocyte, and neuron. could be ruled out.. Immunoreaction to vWF antigen was widespread througout the cytoplasm as Weibel-Palade granule. Immunoreaction to tight junction proteins, i.e. occludin, ZO-1, and ZO-2 was seen at cell contact. In summary, RBMECs isolated and cultured showed morphological, immunohistochemical and electrical characteristics of blood-brain barrier (BBB). The in vitro BBB model can be used in studying characteristics of in vivo BBB.

쥐의 뇌로부터 미세혈관에서 분리하고 배양한 내피세포의 특성을 현미경관찰, 면역염색과 전기저항을 측정해 관찰하였다. 미세혈관 내피세포는 배양 후 $5{\sim}6$일 경에 단층을 형성하였으며 특징적으로 소용돌이치는 모양을 나타냈다. 내피세포 단층의 전기저항은 배양 후 5일 경까지 따라 증가하였고 이후로는 감소하였다. 면역형광염색에서 anti-GFAP, anti-GalC, anti-neurofilament 160/200 kD antibody에 대한 면역반응을 거의 찾아볼 수 없었어 별아교세포, 희소돌기아교세포 및 신경세포에 의한 우려할 만한 오염은 배제할 수 있었다. vWF 항원에 대한 면역반응은 내피세포의 세포질에 Weibel-Palade 과립이 전반적으로 퍼져 있었다. 치밀이음부를 구성하는 occludin, ZO-1, ZO-2에 대한 면역반응은 내피세포의 접촉부위에서 매우 특징적으로 나타나고 있었다. 요약하면 쥐의 뇌 미세혈관 내피세포를 분리, 배양하여 형태학적, 조직면역 화학적 방법과 전기저항을 측정하여 내피세포의 특성을 확인하였다. 이 생체외 혈액뇌장벽 모델은 앞으로 진행될 액뇌장벽의 특징을 규명하고자 하는 시험관내 실험에 유용하게 이용될 수 있을 것이다.

Keywords

References

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