CAJ | 학술논문

目的：建立体外联合肝细胞生长因子（HGF）和表皮生长因子（EGF）培养人胆囊上皮细胞（HGBECs）方法。方法先剥离上皮，采用反复Ⅳ型胶原酶消化配合刮取HGBECs，制备成单细胞悬液进行接种，细胞分别接种于含或不含10ng/mLEGF以及含10ng/mLHGF＋10ng/mLEGF的培养基进行原代培养。倒置显微镜下观察细胞生长的形态变化，细胞计数、噻唑蓝（MTT）法检测HGF＋EGF对HGBECs增殖的影响。结果成功培养HGBECs。添加HGF＋EGF组较添加EGF组HG-BECs增殖速度快且维持时间长；体外存活时间明显延长（19．3±2．5）dvs．（14．2±2．4）d，P＜0．05，细胞活力和细胞形态优于只添加EGF组。结论HGF联合EGF可以明显地促进HGBECs增殖，延长细胞体外存活时间，较长时间稳定细胞形态。
목적：건입체외연합간세포생장인자（HGF）화표피생장인자（EGF）배양인담낭상피세포（HGBECs）방법。방법선박리상피，채용반복Ⅳ형효원매소화배합괄취HGBECs，제비성단세포현액진행접충，세포분별접충우함혹불함10ng/mLEGF이급함10ng/mLHGF＋10ng/mLEGF적배양기진행원대배양。도치현미경하관찰세포생장적형태변화，세포계수、새서람（MTT）법검측HGF＋EGF대HGBECs증식적영향。결과성공배양HGBECs。첨가HGF＋EGF조교첨가EGF조HG-BECs증식속도쾌차유지시간장；체외존활시간명현연장（19．3±2．5）dvs．（14．2±2．4）d，P＜0．05，세포활력화세포형태우우지첨가EGF조。결론HGF연합EGF가이명현지촉진HGBECs증식，연장세포체외존활시간，교장시간은정세포형태。
Objective To establish the method of combined hepatocyte growth factor (HGF) with epidermal growth factor (EGF) cultured human gallbladder epithelial cells(HGBECs) in vitro .Methods The epithelial layer was peeled away from human gallbladder ,epithelial layer were digested with collagenase Ⅳ and scraped repeatedly .HGBECs were isolated and seeded in cell cul-ture plates containing medium supplemented with or without 10 ng/mL EGF or with 10 ng/mL HGF and 10 ng/mL EGF respec-tively .Then the morphologic changes of the cells were observed and taken photos with inverted phase contrast microscope ,and counted number of cells ,MTT assay detected vigor of cells in different groups .Results The number of the HGBECs of the HGF+EGF group was obviously more than the EGF group ,the duration of the HGBECs of the HGF+ EGF group was obviously longer than the EGF group(19 .3 ± 2 .5)d vs .(14 .2 ± 2 .4)d ,P< 0 .05 .And the HGBECs of the group with HGF+ EGF had better cell vigor .Conclusion HGF combines with EGF added to medium can obviously promote the proliferation of HGBECs and prolong the duration and stabilize morphology of HGBECs in vitro .