Fetal bovine serum(FBS) is widely used in cell cultures due to its high stability and easy access. It was also used as a substitute for porcine follicular fluid(PFF) in previous studies. However, FBS components are un...Fetal bovine serum(FBS) is widely used in cell cultures due to its high stability and easy access. It was also used as a substitute for porcine follicular fluid(PFF) in previous studies. However, FBS components are unclear, and the presence of FBS in culture media may introduce a variation from batch to batch. This study aimed to establish an effective method to screen FBS in place of PFF in the culture media for porcine oocytes in vitro. We screened FBS from different sources by using porcine fetal fibroblast cells. The effects of six FBS samples on porcine fetal fibroblast cell growth were tested via frozen cell survival assay, cell clone formation assay, cell growth curve, and cell passage activity assay. The best serum that we called GFBS(heat-inactivated FBS, cat. no. 10500-64;Gibco) showed a similar effect on the maturation and development of porcine oocytes to that of PFF and can be used as a good substitute for PFF. These results suggested that the porcine fetal fibroblast cell culture test can be used as a valuable method to screen FBS for porcine oocyte maturation and embryonic development in vitro.展开更多
BACKGROUND: How to induce endogenous neural stem cells (NSCs) to differentiate into needed neural cell types is a hot spot of current researches. OBJECTIVE: To compare differences between fetal bovine serum and Ch...BACKGROUND: How to induce endogenous neural stem cells (NSCs) to differentiate into needed neural cell types is a hot spot of current researches. OBJECTIVE: To compare differences between fetal bovine serum and Chinese herbal formula Naoluoxintong serum supplementation for inducing proliferation and differentiation in rat embryonic NSCs. DESIGN, TIME AND SETTING: An in vitro, serum pharmacology, comparative, observation study was performed from March to September in 2008 at the Laboratory of Neurodegenerative Diseases, College of Life Science in University of Science and Technology of China, the Key Laboratory Breeding Base of Acupuncture Foundation and Technology in Anhui University of Traditional Chinese Medicine, the Anhui Province Key Laboratory of R & D of Chinese Medicine, and at the Level 3 Laboratory of Molecular Biology of the State Administration of Traditional Chinese Medicine. MATERIALS: The Chinese herbal formula Naoluoxintong was produced by Radix Astragali, Radix Notoginseng, Rhizoma Chuanxiong, Scolopendra at Anhui University of Traditional Chinese Medicine. Mouse anti-rat nestin, gliat fibrillary acidic protein, and galactocerebroside monoclonal antibodies, as well as rabbit anti-neuron-specific enolase polyclonal antibody were produced by Chemicon, Billerica, MA, USA. METHODS: Wistar rats aged 3 months were intragastrically infused with Naoluoxintong. Wistar rat embryonic NSCs (passage 8) were induced to proliferate and differentiate using 10% fetal bovine serum, 10% Naoluoxintong serum, and 10% rat serum. MAIN OUTCOME MEASURES: Phenotypic changes in cultured cells were detected using phase contrast microscopy, and cell proliferation and differentiation were observed using immunofluorescence staining. RESULTS: Proliferation and differentiation of embryonic NSCs was induced by three different types of blood serum. Although the differentiation time course with Nao/uoxintongserum was later than with the other two methods, the differentiated cells were morphologically similar to mature neurons to a greater extent. CONCLUSION: Nao/uoxintong serum supplementation induced differentiation of NSCs into neuronal-like cells and stimulated neuronal maturation.展开更多
The interaction between organic molecules and biomaterial surfaces determines the fate of biomaterials during their service life,which is also the research hotspots in the field of biomaterials.To understand the mecha...The interaction between organic molecules and biomaterial surfaces determines the fate of biomaterials during their service life,which is also the research hotspots in the field of biomaterials.To understand the mechanism of protein interaction with magnesium(Mg)degradation,alloying elements,immersion time,protein concentration and surface conditions have been previously considered for the effect of proteins on Mg degradation.However,fluid flow,as one of the critical factors,drew little attention in this case.In the present study,the effect of bovine serum albumin(BSA)and fetal bovine serum(FBS)on Mg degradation was compared under static and dynamic conditions.The results revealed that both BSA and FBS slightly decreased the degradation rate of Mg in Hanks’balanced salt solution(HBSS)under static immersion due to the protein adsorption and the formation of a Ca/P-rich top layer on Mg surface,whereas under dynamic flow condition the degradation of Mg was significantly accelerated in the presence of BSA or FBS.The reasons seemed to stem from the weakened protein adsorption on Mg surface in this case and the dynamically enhanced interaction between proteins and ions/products in solutions,which largely weaken the combination of the top Ca/P-rich layer with the inner corrosion product layer.These results highlight the importance of testing conditions for Mg characterization in vitro and the synergistic effect between different parameters on Mg degradation.展开更多
Atoh1 overexpression in cochlear epithelium induces new hair cell formation. Use of adenovirus-mediated Atoh1 overexpression has mainly focused on the rat lesser epithelial ridge and induces ectopic hair cell regenera...Atoh1 overexpression in cochlear epithelium induces new hair cell formation. Use of adenovirus-mediated Atoh1 overexpression has mainly focused on the rat lesser epithelial ridge and induces ectopic hair cell regeneration. The sensory region of rat cochlea is difficult to transfect, thus new hair cells are rarely produced in situ in rat cochlear explants. After culturing rat cochleae in medium containing 10% fetal bovine serum, adenovirus successfully infected the sensory region as the width of the supporting cell area was significantly increased. Adenovirus encoding Atoh1 infected the sensory region and induced hair cell formation in situ. Combined application of the Notch inhibitor DAPT and Atoh1 increased the Atoh1 expression level and decreased hes1 and hes5 levels, further promoting hair cell generation. Our results demonstrate that DAPT enhances Atoh1 activity to promote hair cell regeneration in rat cochlear sensory epithelium in vitro.展开更多
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
基金Chongqing Technology Innovation and Application Development Grant,Grant/Award Number:cstc2019jscx-msxm X0394Chongqing Special Fund for Performance Incentive Guide,Grant/Award Number:cqjxjl201709+1 种基金Earmarked Fund for Modern Agro-industry Technology Research System,Grant/Award Number:CARS-36Chongqing Agriculture Development Grant,Grant/Award Number:17406
文摘Fetal bovine serum(FBS) is widely used in cell cultures due to its high stability and easy access. It was also used as a substitute for porcine follicular fluid(PFF) in previous studies. However, FBS components are unclear, and the presence of FBS in culture media may introduce a variation from batch to batch. This study aimed to establish an effective method to screen FBS in place of PFF in the culture media for porcine oocytes in vitro. We screened FBS from different sources by using porcine fetal fibroblast cells. The effects of six FBS samples on porcine fetal fibroblast cell growth were tested via frozen cell survival assay, cell clone formation assay, cell growth curve, and cell passage activity assay. The best serum that we called GFBS(heat-inactivated FBS, cat. no. 10500-64;Gibco) showed a similar effect on the maturation and development of porcine oocytes to that of PFF and can be used as a good substitute for PFF. These results suggested that the porcine fetal fibroblast cell culture test can be used as a valuable method to screen FBS for porcine oocyte maturation and embryonic development in vitro.
基金National Natural Science Foundation of China,No.30873293,30672592Natural Science Foundation of Anhui Province,No.070413125,050430904+1 种基金Dr.Yafang Lü Graduate Research Foundation of Beijing University of Chinese Medicine,No.2004Natural Science Research Fund of Education Department of Anhui Province,No.2006KJ382B
文摘BACKGROUND: How to induce endogenous neural stem cells (NSCs) to differentiate into needed neural cell types is a hot spot of current researches. OBJECTIVE: To compare differences between fetal bovine serum and Chinese herbal formula Naoluoxintong serum supplementation for inducing proliferation and differentiation in rat embryonic NSCs. DESIGN, TIME AND SETTING: An in vitro, serum pharmacology, comparative, observation study was performed from March to September in 2008 at the Laboratory of Neurodegenerative Diseases, College of Life Science in University of Science and Technology of China, the Key Laboratory Breeding Base of Acupuncture Foundation and Technology in Anhui University of Traditional Chinese Medicine, the Anhui Province Key Laboratory of R & D of Chinese Medicine, and at the Level 3 Laboratory of Molecular Biology of the State Administration of Traditional Chinese Medicine. MATERIALS: The Chinese herbal formula Naoluoxintong was produced by Radix Astragali, Radix Notoginseng, Rhizoma Chuanxiong, Scolopendra at Anhui University of Traditional Chinese Medicine. Mouse anti-rat nestin, gliat fibrillary acidic protein, and galactocerebroside monoclonal antibodies, as well as rabbit anti-neuron-specific enolase polyclonal antibody were produced by Chemicon, Billerica, MA, USA. METHODS: Wistar rats aged 3 months were intragastrically infused with Naoluoxintong. Wistar rat embryonic NSCs (passage 8) were induced to proliferate and differentiate using 10% fetal bovine serum, 10% Naoluoxintong serum, and 10% rat serum. MAIN OUTCOME MEASURES: Phenotypic changes in cultured cells were detected using phase contrast microscopy, and cell proliferation and differentiation were observed using immunofluorescence staining. RESULTS: Proliferation and differentiation of embryonic NSCs was induced by three different types of blood serum. Although the differentiation time course with Nao/uoxintongserum was later than with the other two methods, the differentiated cells were morphologically similar to mature neurons to a greater extent. CONCLUSION: Nao/uoxintong serum supplementation induced differentiation of NSCs into neuronal-like cells and stimulated neuronal maturation.
基金the financial supports from China Scholarship Council(CSC,201509350010)Helmholtz Association of German Research Centres.
文摘The interaction between organic molecules and biomaterial surfaces determines the fate of biomaterials during their service life,which is also the research hotspots in the field of biomaterials.To understand the mechanism of protein interaction with magnesium(Mg)degradation,alloying elements,immersion time,protein concentration and surface conditions have been previously considered for the effect of proteins on Mg degradation.However,fluid flow,as one of the critical factors,drew little attention in this case.In the present study,the effect of bovine serum albumin(BSA)and fetal bovine serum(FBS)on Mg degradation was compared under static and dynamic conditions.The results revealed that both BSA and FBS slightly decreased the degradation rate of Mg in Hanks’balanced salt solution(HBSS)under static immersion due to the protein adsorption and the formation of a Ca/P-rich top layer on Mg surface,whereas under dynamic flow condition the degradation of Mg was significantly accelerated in the presence of BSA or FBS.The reasons seemed to stem from the weakened protein adsorption on Mg surface in this case and the dynamically enhanced interaction between proteins and ions/products in solutions,which largely weaken the combination of the top Ca/P-rich layer with the inner corrosion product layer.These results highlight the importance of testing conditions for Mg characterization in vitro and the synergistic effect between different parameters on Mg degradation.
基金supported by the National Natural Science Foundation of China,No.81420108010,81271084,81200740,81371093
文摘Atoh1 overexpression in cochlear epithelium induces new hair cell formation. Use of adenovirus-mediated Atoh1 overexpression has mainly focused on the rat lesser epithelial ridge and induces ectopic hair cell regeneration. The sensory region of rat cochlea is difficult to transfect, thus new hair cells are rarely produced in situ in rat cochlear explants. After culturing rat cochleae in medium containing 10% fetal bovine serum, adenovirus successfully infected the sensory region as the width of the supporting cell area was significantly increased. Adenovirus encoding Atoh1 infected the sensory region and induced hair cell formation in situ. Combined application of the Notch inhibitor DAPT and Atoh1 increased the Atoh1 expression level and decreased hes1 and hes5 levels, further promoting hair cell generation. Our results demonstrate that DAPT enhances Atoh1 activity to promote hair cell regeneration in rat cochlear sensory epithelium in vitro.
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.