CAJ | 학술논문

目的建立微囊化人胃癌细胞株SGC-7901培养模型.方法微胶囊制备仪包裹SGC-7901细胞,培养至21 d,噻唑蓝(MTT)比色法观察隔日细胞的生长活性,生物传感分析仪检测隔日培养上清中葡萄糖和乳酸浓度；选取第7、14、21天的微囊化SGC-7901细胞,苏木素-伊红(HE)染色；免疫细胞化学法分别观察微囊前后SGC-7901细胞5-溴脱氧尿嘧啶核苷(5-BrdU)、细胞核抗原(PCNA)及血管内皮生长因子(VEGF)基因表达.结果 MTT结果示培养至第14天,细胞相对数不再增加,随后细胞相对数略下降；微囊化人胃癌细胞培养上清中葡萄糖含量逐渐降低,乳酸上升；至20d后分别为10、100 mmol/L.免疫细胞化学法检测SGC-7901细胞贴壁培养中表达PCNA和VEGF基因；第7和14天,微囊化人胃癌细胞可见PCNA、BrdU及VEGF基因表达；21 d,PCNA和BrdU主要在细胞团外层表达,内层细胞则表达少或不表达,而VEGF均见表达.结论呈三维立体生长的微囊化胃癌细胞模型生长稳定,相关基因表达稳定.
목적 건립미낭화인위암세포주SGC-7901배양모형.방법 미효낭제비의포과SGC-7901세포,배양지21 d,새서람(MTT)비색법관찰격일세포적생장활성,생물전감분석의검측격일배양상청중포도당화유산농도；선취제7、14、21천적미낭화SGC-7901세포,소목소-이홍(HE)염색；면역세포화학법분별관찰미낭전후SGC-7901세포5-추탈양뇨밀정핵감(5-BrdU)、세포핵항원(PCNA)급혈관내피생장인자(VEGF)기인표체.결과 MTT결과시배양지제14천,세포상대수불재증가,수후세포상대수략하강；미낭화인위암세포배양상청중포도당함량축점강저,유산상승；지20d후분별위10、100 mmol/L.면역세포화학법검측SGC-7901세포첩벽배양중표체PCNA화VEGF기인；제7화14천,미낭화인위암세포가견PCNA、BrdU급VEGF기인표체；21 d,PCNA화BrdU주요재세포단외층표체,내층세포칙표체소혹불표체,이VEGF균견표체.결론 정삼유입체생장적미낭화위암세포모형생장은정,상관기인표체은정.
Objective To establish the cultured model of microencapsulated human gastric carcinoma (HGC) cell line SGC-7901.Methods The SGC-7901 cells were cultured and encapsulated in APA microcapsules using the prepared electrostatic droplet generator for 21 days.The level of proliferation and metabolism of the microencapsulated HGC cells were detected by using methyl thiazol tetrazolium ( MTT),and glucose and lactic acid concentrations were determined in the supernatant.Microencapsulated HGC cells were stained with hematoxylin and eosin (HE),and the expression of bromodeoxyuridine ( BrdU),vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) gene was detected by using immunocytochemistry on the 7th,14th and 21st day.Monolayer cultured cells served as controls.Results On the 14th day,the relative number of the microencapsulated cells was no longer increased and decreased slightly on the next few days.Glucose was consumed and its concentration was decreased to 10 mmol/L,and lactic acid was generated and its concentration was increased to 100 mol/L in microencapsulated HGC cells on the 20th day.The expression of PCNA and VEGF gene could be detected in SGC7901 cells.The expression of BrdU,PCNA and VEGF was still detected in the microencapsulated SGC9701 cells on the 7th and 14th day.On the 21st day,the expression of Brdu and PCNA was found on the outer layer of spheroid,and that of VEGF gene was detected within the whole spheroid.Conclusion The cultured model of microencapsulated HGC SGC9701 had been established,and gene expression was stable.