쥐의 뇌 미세혈관 내피세포를 이용한뇌혈관장벽 내피세포의 배양과 특성
Primary Culture and Characteristics of Blood-Brain BarrierEndothelial Cells from Rat Brain Microvessel
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쥐의 뇌로부터 미세혈관에서 분리하고 배양한 내피세포의 특성을 현미경관찰, 면역염색과 전기저항을 측정해 관찰하였다. 미세혈관 내피세포는 배양 후 5~6일 경에 단층을 형성하였으며 특징적으로 소용돌이치는 모양을 나타냈다. 내피세포 단층의 전기저항은 배양 후 5일경 까지 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 우려 할 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 혈액뇌장벽의 특징을 규명하고자 하는 시험관내 실험에 유용하게 이용될 수 있을 것이다.
Abstract
The characteristics of primary cultured rat brain microvessel endothelial cells(RBMECs) were studied usingRBMECs formed a monolayer by 5~ 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-neuron. could be ruled out.. Immunoreaction to vWF antigen was widespread througout the cytoplasm asWeibel-Palade granule. Immunoreaction to tight junction proteins, i.e. occludin, ZO-1, and ZO-2 was seen atcell contact. In summary, RBMECs isolated and cultured showed morphological, immunohistochemical and-brain barrier (BBB). The in vitro BBB model can be used in studyingcharacteristics of in vivo BBB.
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