中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1925-1928
,共4页
沈剑虹%杨甫%姚麒%顾志恺%周非
瀋劍虹%楊甫%姚麒%顧誌愷%週非
침검홍%양보%요기%고지개%주비
第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因%抑癌基因%神经干细胞%壳聚糖%神经组织工程
第10號染色體上缺失與張力蛋白同源的燐痠酯酶基因%抑癌基因%神經榦細胞%殼聚糖%神經組織工程
제10호염색체상결실여장력단백동원적린산지매기인%억암기인%신경간세포%각취당%신경조직공정
Phosphatase and tensin homologue deleted on chromosometen%Tumor suppressor%Neural stem cells%Chitosan%Nerve tissue engineering
目的 观察神经干细胞在吸附第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因(PTEN)抑制剂的壳聚糖支架上的生长分化.方法 神经干细胞分壳聚糖组(培养孔内置入壳聚糖支架)和对照组,于1、3、7d行细胞计数试剂盒(CCK-8)实验,比较两组细胞的增殖.制备吸附PTEN抑制剂bpV(pic)的壳聚糖支架,以空支架为对照,与神经干细胞共培养.培养7d时取壳聚糖支架冰冻切片,免疫荧光染色鉴定其分化.结果 CCK-8结果显示,各时间点壳聚糖组(0.267±0.012、0.444±0.019、0.787-±0.031)和对照组(0.237±0.022、0.467 ±0.024、0.772±0.021)的吸光度值差异无统计学意义(P＞0.05).免疫荧光染色显示,与空支架组比较,bpV支架组抗胶质纤维酸性蛋白(GFAP)阳性细胞比例[(73.2±9.1)％比(85.2±9.5)％]较低,而抗β-微管蛋白(β-tubulin)Ⅲ阳性细胞比例[(24.6±8.9)％比(11.4±7.2)％]较高(P＜0.05),每视野GFAP和β-Tubulin Ⅲ阳性细胞数在bpv支架组(11.4±2.0、4.1±1.7)都多于空支架组(8.8±2.3、1.3±0.8,P＜0.05);两组均少见有抗受体相互作用蛋白(RIP)阳性细胞.结论 壳聚糖支架对神经干细胞有良好的相容性,吸附PTEN抑制剂的壳聚糖支架能促进神经干细胞向支架迁移并向神经元分化.
目的 觀察神經榦細胞在吸附第10號染色體上缺失與張力蛋白同源的燐痠酯酶基因(PTEN)抑製劑的殼聚糖支架上的生長分化.方法 神經榦細胞分殼聚糖組(培養孔內置入殼聚糖支架)和對照組,于1、3、7d行細胞計數試劑盒(CCK-8)實驗,比較兩組細胞的增殖.製備吸附PTEN抑製劑bpV(pic)的殼聚糖支架,以空支架為對照,與神經榦細胞共培養.培養7d時取殼聚糖支架冰凍切片,免疫熒光染色鑒定其分化.結果 CCK-8結果顯示,各時間點殼聚糖組(0.267±0.012、0.444±0.019、0.787-±0.031)和對照組(0.237±0.022、0.467 ±0.024、0.772±0.021)的吸光度值差異無統計學意義(P＞0.05).免疫熒光染色顯示,與空支架組比較,bpV支架組抗膠質纖維痠性蛋白(GFAP)暘性細胞比例[(73.2±9.1)％比(85.2±9.5)％]較低,而抗β-微管蛋白(β-tubulin)Ⅲ暘性細胞比例[(24.6±8.9)％比(11.4±7.2)％]較高(P＜0.05),每視野GFAP和β-Tubulin Ⅲ暘性細胞數在bpv支架組(11.4±2.0、4.1±1.7)都多于空支架組(8.8±2.3、1.3±0.8,P＜0.05);兩組均少見有抗受體相互作用蛋白(RIP)暘性細胞.結論 殼聚糖支架對神經榦細胞有良好的相容性,吸附PTEN抑製劑的殼聚糖支架能促進神經榦細胞嚮支架遷移併嚮神經元分化.
목적 관찰신경간세포재흡부제10호염색체상결실여장력단백동원적린산지매기인(PTEN)억제제적각취당지가상적생장분화.방법 신경간세포분각취당조(배양공내치입각취당지가)화대조조,우1、3、7d행세포계수시제합(CCK-8)실험,비교량조세포적증식.제비흡부PTEN억제제bpV(pic)적각취당지가,이공지가위대조,여신경간세포공배양.배양7d시취각취당지가빙동절편,면역형광염색감정기분화.결과 CCK-8결과현시,각시간점각취당조(0.267±0.012、0.444±0.019、0.787-±0.031)화대조조(0.237±0.022、0.467 ±0.024、0.772±0.021)적흡광도치차이무통계학의의(P＞0.05).면역형광염색현시,여공지가조비교,bpV지가조항효질섬유산성단백(GFAP)양성세포비례[(73.2±9.1)％비(85.2±9.5)％]교저,이항β-미관단백(β-tubulin)Ⅲ양성세포비례[(24.6±8.9)％비(11.4±7.2)％]교고(P＜0.05),매시야GFAP화β-Tubulin Ⅲ양성세포수재bpv지가조(11.4±2.0、4.1±1.7)도다우공지가조(8.8±2.3、1.3±0.8,P＜0.05);량조균소견유항수체상호작용단백(RIP)양성세포.결론 각취당지가대신경간세포유량호적상용성,흡부PTEN억제제적각취당지가능촉진신경간세포향지가천이병향신경원분화.
Objective To investigate the growth and differentiation of neural stem cells cultured on chitosan scaffolds combined with phosphatase and tensin homologue deleted on chromosometen (PTEN) inhibitor.Methods Neural stem cells (NSCs) were divided into chitosan group (with chitosan scaffolds in culture hole) and control group,and cell counting kit-8 (CCK-8) test was carried out to compare cell proliferation of the two groups on 1,3 and 7 d.Then bpV (pic) was absorbed to chitosan scaffolds and cultured with 2 groups of NSCs (bpV scaffolds group and non-loaded scaffolds group).Chitosan scaffolds were frozen sectioned and immunofluorescence staining was used to detect the differentiation of NSCs.Results CCK-8 test showed no significant difference of absorbance existing between chitosan group (0.267 ±0.012,0.444 ±0.019,0.787 ±0.031) and control group (0.237 ±0.022,0.467 ±0.024,0.772 ± 0.021) at each time point.Immunofluorescence staining showed that lower glial fibrillary acidic protein (GFAP) + cell proportion [(73.2 ±9.1)％ vs.(85.2 ±9.5)％] and higher β-tubulin Ⅲ + cell proportion [(24.6 ± 8.9) ％ vs.(11.4 ± 7.2) ％)] were detected in bpv scaffolds than in non-loaded scaffolds.GFAP + cells and β-tubulin Ⅲ + cells were both more in bpv scaffolds (11.4 ± 2.0 and 4.1 ± 1.7 respectively) than in non-loaded scaffolds (8.8 ±2.3 and 1.3 ±0.8 respectively),but receptor interacting protein (RIP) + cells were rare in the two groups.Conclusion This kind of chitosan scaffolds has good biocompatibility to NSCs.Chitosan scaffolds combined with PTEN inhibitor can promote NSCs to migrate towards the scaffolds and differentiate to neurons.
