CAJ | 학술논문

目的：：探讨小鼠牙囊细胞的体外培养方法并观察其形态特征。方法：体视显微镜下分离出生3~5d仔鼠牙胚,用I型胶原酶消化后分离牙囊组织,采用酶消化-组织块法和二次酶消化法进行牙囊细胞的原代及传代培养,倒置显微镜下进行细胞形态学观察。结果：酶消化-组织块法经4~5 d,部分组织块周围见细胞呈放射状爬出,2周后细胞集落仍难以汇合,无法传代。二次酶消化法第2天可见散在细胞集落形成,4~5d细胞接触汇合。传代发现,多角形细胞消失,第2代后梭形细胞占大多数,第3代细胞生长速度最快,第4代细胞增殖速度减慢,第5代后细胞趋于静止；随着传代次数的增加胞体逐渐增大,可出现一些树突形细胞。结论：采用二次酶消化法进行小鼠牙囊细胞原代培养可在短时间内获得较多的细胞,操作简便且成功率高,细胞形态随培养时间而变化多样。
목적：：탐토소서아낭세포적체외배양방법병관찰기형태특정。방법：체시현미경하분리출생3~5d자서아배,용I형효원매소화후분리아낭조직,채용매소화-조직괴법화이차매소화법진행아낭세포적원대급전대배양,도치현미경하진행세포형태학관찰。결과：매소화-조직괴법경4~5 d,부분조직괴주위견세포정방사상파출,2주후세포집락잉난이회합,무법전대。이차매소화법제2천가견산재세포집락형성,4~5d세포접촉회합。전대발현,다각형세포소실,제2대후사형세포점대다수,제3대세포생장속도최쾌,제4대세포증식속도감만,제5대후세포추우정지；수착전대차수적증가포체축점증대,가출현일사수돌형세포。결론：채용이차매소화법진행소서아낭세포원대배양가재단시간내획득교다적세포,조작간편차성공솔고,세포형태수배양시간이변화다양。
AIM:To explore the culture method of mouse dental follicle cells ( DFCs) in vitro and to observe their morphological characteristics. METHODS:Dental germs from 3-5 days postnatal mice were digested with type I collagenase and the dental follicle tissues were dissected by enzyme isolation-tissue explant and twice-enzyme isola-tion. The morphology of the cultured DFCs was observed under inverted microscope. RESULTS:At the 4 th-5 th day in vitro, several cells were extended from part of the tissue explants and were hardly to fuse even after 2 weeks′culture. The method of enzyme isolation-tissue explant was failed because of unable to passage and amplify the cells while the method of twice-enzyme isolation got a success rate of about 85%. Scattered groups of cells were seen at the second day and they fused at the 4 th-5 th day in vitro by way of twice-enzyme isolation. Polygonal and spindle cell populations co-existed in primary culture. Almost all polygonal cells disappeared in subculture, and spindle cells became the majority of cell population. The cells of third passage grew fastest, then slowed down and tended to stop in the fourth and fifth pas-sage, respectively. After several times of passage, the cells became larger and polymorphic in shape, some dendrite-like cells were observed. CONCLUSION:The method of twice-enzyme isolation may havest considerable cells in a short time with a high success rate. Morphology of DFCs is changed with the culture time and is multiple.