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Differentiation of mesenchymal stem cells into neuronal cells on fetal bovine acellular dermal matrix as a tissue engineered nerve scaffold 被引量:4
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作者 Yuping Feng Jiao Wang +5 位作者 Shixin Ling Zhuo Li Mingsheng Li Qiongyi Li Zongren Ma Sijiu Yu 《Neural Regeneration Research》 SCIE CAS CSCD 2014年第22期1968-1978,共11页
The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural di... The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined speciifc neu-ronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuro-nal-speciifc proteins, includingβIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differen-tiation medium differentiated into a multilayered neural network-like structure with long nerve ifbers that was composed of several parallel microifbers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sec-tioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve. 展开更多
关键词 nerve regeneration peripheral nerve defects fetal bovine acellular dermal matrix biological scaffold bone marrow mesenchymal stem cells neuronal differentiation neurons tissue engineered nerve neural regeneration
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Screening high-quality fetal bovine serum for porcine oocyte maturation in vitro 被引量:3
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作者 Xueqing Liu Qiaoli Lang +8 位作者 Meng Wu Xiaoyan You Qiling He Ling Luo Zijia Liu Puying Xiao Nan Huang Xi Yang Liangpeng Ge 《Animal Models and Experimental Medicine》 CSCD 2019年第4期334-339,共6页
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
关键词 fetal bovine serum maturation rate porcine oocytes
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Fetal bovine serum versus Chinese herbal formula Naoluoxintong serum supplementation for proliferation and differentiation of rat embryonic neural stem cells
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作者 Wei Tang Jian Wang +6 位作者 Youwen Wang Chaomin Ni Yenong Chen Zhaoliang Tang Lihua Yu Xiaomin Li Jianpeng Hu 《Neural Regeneration Research》 SCIE CAS CSCD 2010年第14期1061-1065,共5页
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
关键词 neural stem cells DIFFERENTIATION PROLIFERATION Chinese herbal formula Nao/uoxintong drug serum fetal bovine serum neural regeneration
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Evaluation of the major royal jelly proteins as an alternative to fetal bovine serum in culturing human cell lines
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作者 Di CHEN Xiao-xuan XIN +2 位作者 Hao-cheng QIAN Zhang-yin YU Li-rong SHEN 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2016年第6期476-483,共8页
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
关键词 Major royal jelly proteins (MRJPs) Cell culture ALTERNATIVE fetal bovine serum (FBS) CYTOKINE
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Comparing successful gene knock-in efficiencies of CRISPR/Cas9 with ZFNs and TALENs gene editing systems in bovine and dairy goat fetal fibroblasts 被引量:11
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作者 LIU Hui LIU Chang +5 位作者 ZHAO Yu-hang HAN Xue-jie ZHOU Zheng-wei WANG Chen LI Rong-feng LI Xue-ling 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第2期406-414,共9页
This study aimed to compare the efficiencies of clustered regulatory interspaced short palindromic repeat(CRISPR)/Cas9-mediated gene knock-ins with zinc finger nucleases(ZFNs) and transcription activator-like effe... This study aimed to compare the efficiencies of clustered regulatory interspaced short palindromic repeat(CRISPR)/Cas9-mediated gene knock-ins with zinc finger nucleases(ZFNs) and transcription activator-like effector nucleases(TALENs) in bovine and dairy goat fetal fibroblasts. To test the knock-in efficiency, a set of ZFNs and CRISPR/Cas9 plasmids were designed to edit the bovine myostatin(MSTN) gene at exon 2, while a set of TALENs and CRISPR/Cas9 plasmids were designed for editing the dairy goat β-casein gene at exon 2. Donor plasmids utilizing the ZFNs, TALENs, and CRISPR/Cas9 cutting sites were constructed in theGFP-PGK-Neo R plasmid background, including a 5′ and 3′ homologous arm flanking the genes humanized Fat-1(h Fat-1) or enhanced green fluorescent protein(eGFP). Subsequently, the ZFNs, TALENs, or CRISPR/Cas9 and thehFat-1 or eGFP plasmids were co-transfected by electroporation into bovine and dairy goat fetal fibroblasts. After G418(Geneticin) selection, single cells were obtained by mouth pipetting, flow cytometry or a cell shove. The gene knock-in events were screened by PCR across the homologous arms. The results showed that in bovine fetal fibrobalsts, the efficiencies of ZFNs-mediated eGFP andhFat-1 gene knock-ins were 13.68 and 0%, respectively. The efficiencies of CRISPR/Cas9-mediated eGFP andhFat-1 gene knock-ins were 77.02 and 79.01%, respectively. The eGFP gene knock-in efficiency using CRISPR/Cas9 was about 5.6 times higher than when using the ZFNs gene editing system. Additionally, thehFat-1 gene knock-in was only obtained when using the CRISPR/Cas9 system. The difference of knockin efficiencies between the ZFNs and CRISPR/Cas9 systems were extremely significant(P〈0.01). In the dairy goat fetal fibroblasts, the efficiencies of TALENs-mediated eGFP andhFat-1 gene knock-ins were 32.35 and 26.47%, respectively. Theefficiencies of eGFP and hFat-1 gene knock-ins using CRISPR/Cas9 were 70.37 and 74.29%, respectively. The knock-in efficiencies difference between the TALENs and CRISPR/Cas9 systems were extremely significant(P〈0.01). This study demonstrated that CRISPR/Cas9 was more efficient at gene knock-ins in domesticated animal cells than ZFNs and TALENs. The CRISPR/Cas9 technology offers a new era of precise gene editing in domesticated animal cell lines. 展开更多
关键词 myostatin(MSTN) β-casein(CSN2) bovine fetal fibroblasts CRISPR/Cas9 dairy goat fetal fibroblasts eGFP hFat-1 knock-in mutation efficiency TALENs ZFNs
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Effects of proteins on magnesium degradation-static vs.dynamic conditions
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作者 Ruiqing Hou Frank Feyerabend +2 位作者 Heike Helmholz Vasil M.Garamus Regine Willumeit-Römer 《Journal of Magnesium and Alloys》 SCIE EI CAS CSCD 2023年第4期1332-1342,共11页
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
关键词 FLOW bovine serum albumin fetal bovine serum Protein adsorption
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Notch pathway inhibitor DAPT enhances Atoh1 activity to generate new hair cells in situ in rat cochleae 被引量:6
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作者 Wen-wei Luo Zhao Han +3 位作者 Dong-dong Ren Xin-wei Wang Fang-lu Chi Juan-mei Yang 《Neural Regeneration Research》 SCIE CAS CSCD 2017年第12期2092-2099,共8页
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
关键词 nerve regeneration Atoh 1 DAPT. transdifferentiation gamma secretase inhibitor COCHLEA sensory epithelium fetal bovine serum hair cell supporting cell hair cell regeneration neural regeneration
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