中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
9期
2074-2076
,共3页
周强%漆松涛%向伟%王海%张立%李宏%刘亚伟%陆云涛
週彊%漆鬆濤%嚮偉%王海%張立%李宏%劉亞偉%陸雲濤
주강%칠송도%향위%왕해%장립%리굉%류아위%륙운도
胶质母细胞瘤%原代培养%单克隆%有限稀释法
膠質母細胞瘤%原代培養%單剋隆%有限稀釋法
효질모세포류%원대배양%단극륭%유한희석법
Glioblastoma%Primary culture%Single cell clone%Limited dilution method
目的 建立胶质母细胞瘤原代细胞的单克隆株并观察记录不同单克隆株间细胞形态和增殖能力的差异.方法 胶质母细胞瘤组织原代培养后进行纯化、单克隆培养、鉴定;定期观察单克隆细胞株的生长状况和形态特征,并比较其增殖能力的差异.结果 成功建立了23株胶质母细胞瘤细胞单克隆株,根据单克隆株形态不同,将其分为长伪足型和短伪足型,长伪足型伪足长(142.50±33.65) μm,短伪足型伪足长(31.33 ±5.72) μm,两者伪足长短差异有统计学意义(P＜0.01);同时,细胞增殖实验证明长伪足型单克隆细胞株增殖能力弱于短伪足型单克隆细胞株,细胞计数试剂盒(CCK--8)实验显示长伪足型单克隆株第10天吸光度值为0.85 ±0.13,短伪足型单克隆株吸光度值为1.31 ±0.20,两者差异有统计学意义(P＜0.01).结论 不同单克隆细胞株形态和增殖能力的差异印证了肿瘤细胞的异质性.
目的 建立膠質母細胞瘤原代細胞的單剋隆株併觀察記錄不同單剋隆株間細胞形態和增殖能力的差異.方法 膠質母細胞瘤組織原代培養後進行純化、單剋隆培養、鑒定;定期觀察單剋隆細胞株的生長狀況和形態特徵,併比較其增殖能力的差異.結果 成功建立瞭23株膠質母細胞瘤細胞單剋隆株,根據單剋隆株形態不同,將其分為長偽足型和短偽足型,長偽足型偽足長(142.50±33.65) μm,短偽足型偽足長(31.33 ±5.72) μm,兩者偽足長短差異有統計學意義(P＜0.01);同時,細胞增殖實驗證明長偽足型單剋隆細胞株增殖能力弱于短偽足型單剋隆細胞株,細胞計數試劑盒(CCK--8)實驗顯示長偽足型單剋隆株第10天吸光度值為0.85 ±0.13,短偽足型單剋隆株吸光度值為1.31 ±0.20,兩者差異有統計學意義(P＜0.01).結論 不同單剋隆細胞株形態和增殖能力的差異印證瞭腫瘤細胞的異質性.
목적 건립효질모세포류원대세포적단극륭주병관찰기록불동단극륭주간세포형태화증식능력적차이.방법 효질모세포류조직원대배양후진행순화、단극륭배양、감정;정기관찰단극륭세포주적생장상황화형태특정,병비교기증식능력적차이.결과 성공건립료23주효질모세포류세포단극륭주,근거단극륭주형태불동,장기분위장위족형화단위족형,장위족형위족장(142.50±33.65) μm,단위족형위족장(31.33 ±5.72) μm,량자위족장단차이유통계학의의(P＜0.01);동시,세포증식실험증명장위족형단극륭세포주증식능력약우단위족형단극륭세포주,세포계수시제합(CCK--8)실험현시장위족형단극륭주제10천흡광도치위0.85 ±0.13,단위족형단극륭주흡광도치위1.31 ±0.20,량자차이유통계학의의(P＜0.01).결론 불동단극륭세포주형태화증식능력적차이인증료종류세포적이질성.
Objective To establish single cell colony from human glioblastoma multiforme (GBM) and observe corresponding morphological characteristics and proliferation capacity.Methods GBM cells were acquired by enzymatic digestion culture method and further purified for single clone culture.Subsequently,these glial fibrillary acidic protein (GFAP) positive colonies were indentified for research.At last,cell growth and morphological characteristics of single cell colony were recorded.Results We successfully establish 23 single cell colonies and divide them into short pseudopod type and long pseudopod type according to their morphological diversity,the length of pseudopod is (142.50 ± 33.65) μm in long pseudopod group and (31.33 ± 5.72) μm in short pseudopod group respectively,length of these two type cell are significantly different (P ＜ 0.01).In addition,cell counting kit-8 (CCK-8) assay indicate short pseudopod type cell exhibit significant higher proliferation capacity than long pseudopod type cell.The OD value at 10th day is 0.85 ±0.13 in long pseudopod group and 1.31 ±0.20 in short pseudopod group respectively (P ＜ 0.01).Conclusion Diversity of morphology and proliferation capacity among cell colonies partially demonstrate existing of intratumor heterogeneity.
