Royal jelly （R J） is a well-known bioactive substance. It contains large amounts of major royal jelly proteins （MRJPs）, which express growth-factor-like activity in several animal and human cell lines. However, th...Royal jelly （R J） is a well-known bioactive substance. It contains large amounts of major royal jelly proteins （MRJPs）, which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum （FBS） in different types of human cell culture, the prolif- eration of the complex serum with different ratios of MRJPs/FBS （M/F） was evaluated on five cell lines： 293T, HFL-I, 231, HCT116, and Changliver using MTT （3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2H-tetrazolium bromide） assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor （EGF） and insulin-transferrin-selenium （ITS）, pro- moted proliferations of Changliver, 293T, HCT116, and H FL-I by 18.73%-56.19% （P〈0.01）. Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.展开更多
目的通过比较含有不同胎牛血清浓度的心肌球培养基分离培养大鼠心肌干细胞(CSCs)的效果,探讨最优的心肌干细胞血清培养浓度。方法选取15只健康新生Wistar大鼠(出生1天龄),无菌条件下取出心脏,经胰酶和Ⅱ型胶原酶反复消化后,种植于培养瓶...目的通过比较含有不同胎牛血清浓度的心肌球培养基分离培养大鼠心肌干细胞(CSCs)的效果,探讨最优的心肌干细胞血清培养浓度。方法选取15只健康新生Wistar大鼠(出生1天龄),无菌条件下取出心脏,经胰酶和Ⅱ型胶原酶反复消化后,种植于培养瓶中,应用含20%胎牛血清的完全培养基培养组织块源性心脏祖细胞(CPCs),将所得的培养外植物源性细胞(EDCs)随机分为A、B、C 3组,各组5瓶细胞(25 m L塑料培养瓶),分别应用含10%、11%、15%不同浓度胎牛血清的心肌球生长培养基培养,培养至第三代CSCs时行细胞表面抗原C-kit染色,鉴定C-kit阳性CSCs占所获得细胞的比率,比较所得的细胞数量,生长周期和生长曲线。结果从新生大鼠心肌组织中成功分离培养出CSCs,C-kit阳性率87%。培养后所得细胞数量以细胞倍增水平(PDL)表示,各组细胞PDL呈匀速递增,PDL水平和其递增速率(生长曲线所示)比较:B组>C组>A组。应用含11%胎牛血清的心肌球生长培养基培养EDCs获得的CSCs,与其他两组相比,具有生长周期短(P<0.05),生长数量多的特点(P<0.05)。结论含11%胎牛血清浓度是心肌球生长培养基培养原代大鼠心肌干细胞的最优血清培养浓度。展开更多
目的探讨人AB血浆培养外周血造血干/祖细胞(HSCs/HPCs)向成熟红细胞分化的可行性。方法将外周血HSCs/HPCs分别在添加5%胎牛血清(FBS)(FBS组)、3%FBS+2%人AB血浆(FBS+AB血浆组)和8%人AB血浆(AB血浆组)中的培养条件下诱导分化为成熟红细胞...目的探讨人AB血浆培养外周血造血干/祖细胞(HSCs/HPCs)向成熟红细胞分化的可行性。方法将外周血HSCs/HPCs分别在添加5%胎牛血清(FBS)(FBS组)、3%FBS+2%人AB血浆(FBS+AB血浆组)和8%人AB血浆(AB血浆组)中的培养条件下诱导分化为成熟红细胞,按照1×10;/mL密度接种在24孔培养板中,2 mL/孔,观察3种培养条件下细胞生长增殖曲线和细胞形态学(细胞涂片,May-Giemsa染色,显微镜下观察细胞形态及细胞核变化)变化情况,流式细胞仪检测细胞表面红系终末分化标志分子糖化血红蛋白A(GPA)、带3蛋白(Band3)和整合素α4(α4-integrin)表达情况,以及红系晚期细胞脱核情况;比较3种培养条件对外周血HSCs/HPCs向成熟红细胞分化的影响。结果 AB血浆组、FBS组、FBS+AB血浆组的细胞生长增殖倍数分别为2 573±116 vs 2 514±246 vs 2 539±119(P>0.05);3组细胞形态学变化相似:随着培养时间延长,HSCs/HPCs从原幼红细胞向嗜碱性幼红细胞、多染幼红细胞、正染幼红细胞分化,至21 d时,几乎全部分化为脱核的红细胞;流式检测红系终末分化及脱核:血型糖蛋白A(GPA)表达及脱核率(%)分别为97.17±1.91 vs 94.95±1.61 vs 96.15±1.38、85.1±3.26 vs 86.93±5.96 vs 86.5±3.36(P>0.05).结论人AB血浆体外培养外周HSCs/HPCs向成熟红细胞分化实验达到了与胎牛血清培养相似的结果。展开更多
基金Project supported by the National Natural Science Foundation of China(No.31271848)the Important Scientific&Technical Innovation Project of Hangzhou(No.20131812A25)the Foundation of Fuli Institute of Food Science of Zhejiang University(No.KY201404),China
文摘Royal jelly （R J） is a well-known bioactive substance. It contains large amounts of major royal jelly proteins （MRJPs）, which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum （FBS） in different types of human cell culture, the prolif- eration of the complex serum with different ratios of MRJPs/FBS （M/F） was evaluated on five cell lines： 293T, HFL-I, 231, HCT116, and Changliver using MTT （3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2H-tetrazolium bromide） assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor （EGF） and insulin-transferrin-selenium （ITS）, pro- moted proliferations of Changliver, 293T, HCT116, and H FL-I by 18.73%-56.19% （P〈0.01）. Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.
文摘目的通过比较含有不同胎牛血清浓度的心肌球培养基分离培养大鼠心肌干细胞(CSCs)的效果,探讨最优的心肌干细胞血清培养浓度。方法选取15只健康新生Wistar大鼠(出生1天龄),无菌条件下取出心脏,经胰酶和Ⅱ型胶原酶反复消化后,种植于培养瓶中,应用含20%胎牛血清的完全培养基培养组织块源性心脏祖细胞(CPCs),将所得的培养外植物源性细胞(EDCs)随机分为A、B、C 3组,各组5瓶细胞(25 m L塑料培养瓶),分别应用含10%、11%、15%不同浓度胎牛血清的心肌球生长培养基培养,培养至第三代CSCs时行细胞表面抗原C-kit染色,鉴定C-kit阳性CSCs占所获得细胞的比率,比较所得的细胞数量,生长周期和生长曲线。结果从新生大鼠心肌组织中成功分离培养出CSCs,C-kit阳性率87%。培养后所得细胞数量以细胞倍增水平(PDL)表示,各组细胞PDL呈匀速递增,PDL水平和其递增速率(生长曲线所示)比较:B组>C组>A组。应用含11%胎牛血清的心肌球生长培养基培养EDCs获得的CSCs,与其他两组相比,具有生长周期短(P<0.05),生长数量多的特点(P<0.05)。结论含11%胎牛血清浓度是心肌球生长培养基培养原代大鼠心肌干细胞的最优血清培养浓度。
文摘目的探讨人AB血浆培养外周血造血干/祖细胞(HSCs/HPCs)向成熟红细胞分化的可行性。方法将外周血HSCs/HPCs分别在添加5%胎牛血清(FBS)(FBS组)、3%FBS+2%人AB血浆(FBS+AB血浆组)和8%人AB血浆(AB血浆组)中的培养条件下诱导分化为成熟红细胞,按照1×10;/mL密度接种在24孔培养板中,2 mL/孔,观察3种培养条件下细胞生长增殖曲线和细胞形态学(细胞涂片,May-Giemsa染色,显微镜下观察细胞形态及细胞核变化)变化情况,流式细胞仪检测细胞表面红系终末分化标志分子糖化血红蛋白A(GPA)、带3蛋白(Band3)和整合素α4(α4-integrin)表达情况,以及红系晚期细胞脱核情况;比较3种培养条件对外周血HSCs/HPCs向成熟红细胞分化的影响。结果 AB血浆组、FBS组、FBS+AB血浆组的细胞生长增殖倍数分别为2 573±116 vs 2 514±246 vs 2 539±119(P>0.05);3组细胞形态学变化相似:随着培养时间延长,HSCs/HPCs从原幼红细胞向嗜碱性幼红细胞、多染幼红细胞、正染幼红细胞分化,至21 d时,几乎全部分化为脱核的红细胞;流式检测红系终末分化及脱核:血型糖蛋白A(GPA)表达及脱核率(%)分别为97.17±1.91 vs 94.95±1.61 vs 96.15±1.38、85.1±3.26 vs 86.93±5.96 vs 86.5±3.36(P>0.05).结论人AB血浆体外培养外周HSCs/HPCs向成熟红细胞分化实验达到了与胎牛血清培养相似的结果。
