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Hierarchically Inverse Opal Porous Scaffolds from Droplet Microfluidics for Biomimetic 3D cell Co-culture

Engineering 2021年第12期7卷 1778-1785页

作者： Changmin Shao Yuxiao Liu Junjie Chi Fangfu Ye Yuanjin Zhao Department of Rheumatology and Immunology Institute of Translational MedicineAffiliated Drum Tower HospitalMedical School of Nanjing UniversityNanjing 210008China Oujiang Laboratory(Zhejiang Lab for Regenerative Medicine Vision and Brain Health)Wenzhou 325038China Wenzhou Institute University of Chinese Academy of SciencesWenzhou 325016China State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Beijing National Laboratory for Condensed Matter Physics Institute of PhysicsChinese Academy of SciencesBeijing 100190China

With the advantages of better mimicking the specificity of natural tissues,three-dimensional(3D)Cell culture plays a major role in drug development,toxicity testing,and tissue engineering.However,existing scaffolds or microcarriers for 3D Cell culture are often limited in size and show suboptimal performance in simulating the vascular complexes of living organisms.Therefore,we present a novel hierarchically inverse opal porous scaffold made via a simple microfluidic approach for promoting 3D cell co-culture techniques.The designed scaffold is constructed using a combined concept involving an emulsion droplet template and inert polymer polymerization.This work demonstrates that the resultant scaffolds ensure a sufficient supply of nutrients during Cell culture,so as to achieve large-volume Cell culture.In addition,by serially planting different cells in the scaffold,a 3D co-culture system of endothelial-cellencapsulated hepatocytes can be developed for constructing certain functional tissues.It is also demonstrated that the use of the proposed scaffold for a co-culture system helps hepatocytes to maintain specific in vivo functions.These hierarchically inverse opal scaffolds lay the foundation for 3D Cell culture and even the construction of biomimetic tissues.

关键词： Microfluidics Inverse opal Cell culture Droplet Biomaterial

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Effects of substance P on growth of fibroblast-like cells derived from bile duct： an in vitro Cell culture study

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Chinese Medical Journal 2014年第17期127卷 3121-3126页

作者： Tian Yuanhu Yang Guangyun Zhang Xiaoqing Shen Wei Dong Jiahong Xu Zhi Department of Hepatobiliary Surgery Chinese People's Liberation Army General Hospital Beijing 100853 China Department of General Surgery Peking University Third Hospital Beijing 100191 China

Background The possible role of substance P （SP） during wound healing has been the primary research focus in recent years,but its effect on the healing process after bile duct injury is little understood.This study aimed to investigate the effects of SP on growth of fibroblast-like cells derived from rabbit bile duct.Methods Fibroblast-like cells derived from rabbit bile duct were identified and divided randomly into control and experimental groups.SP-treated cells at different concentrations of 10^-9-10^-5 mol/L and control group were incubated,respectively,for 48 hours.After incubating,the effects of SP on cell proliferation were assessed by cell counts and MTT test.Apoptosis rate （AR） of cells was measured by flow cytometry.Results cultured rabbit bile duct cells were fibroblast-like in morphology,and these cells were stained positively for vimentin and negatively for desmin.After SP was added to nonconfluent cells for 48 hours,cell numbers were significantly increased in experimental groups than in controls （P 〈0.05）.The maximum stimulation of cell proliferation was achieved at SP of 10^-5 mol/L.Bile duct fibroblast-like cells in the SP group showed a higher proliferating activity and lower AR than those in the control group or in the SP ＋ Spantide group （P 〈0.05）.Spantide partly inhibited the effects of SP on fibroblastlike cells.Examination under transmission electron microscopy revealed rough endoplasmic reticulum and prominent Golgi complexes after SP treatment.Conclusions SP has a growth regulatory property on cultivated bile duct fibroblast-like cells in vitro,suggesting that SP may involve in wound healing after bile duct injury by promoting wound fibroblast proliferation and inhibiting apoptosis and participate in pathological scar formation.

关键词： substance P bile duct fibroblast Cell culture cell proliferation

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Effect of elicitors, precursors and metabolic inhibitors on paclitaxel production by Taxus cuspidata Cell culture

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Journal of Forestry Research 2016年第6期27卷 1257-1263页

作者： Shujie Wang Chun Li Hujun Wang Xiangmei Zhong Jing Zhao Yueying Tong Yajun Zhou College of Biological and Agricultural Engineering Jilin University Key Laboratory of Bionic Engineering (Ministry of Education) Jilin University College of Food Science and Engineering Jilin University

In this study, using Taxus cuspidata as a raw material, we obtained stable high-yielding cell lines by subculturing and quantified paclitaxel content using ultra- sonic extraction combined with TLC-UV spectrophotom- etry. In single factor and multiple factors tests to optimize design and study the effects of elicitors, precursors, and metabolic inhibitors on paclitaxel production by Taxus cuspidata cells, paclitaxel production reached 4.32 mg/L when 100 μmol/L methyl jasmonate, 20 mg/L salicylic acid, 400 mg/L phenylalanine and 2 mg/L gibberellin （GA3） were added to the culture medium of suspension cells. When adding metabolic adjustment factors on the 7th day of culture, extra- and intracellular paclitaxel production was the highest at 4.855 mg/L, paclitaxel release rate was 10.48 %, fresh mass and paclitaxel production of cell increased, respectively, by 6.08 and 11.57 %. By controlling the anabolism of paclitaxel, paclitaxel yield was significantly improved.

关键词： Elicitor Precursor Metabolic inhibitor Paclitaxel Cell culture Taxus cuspidata

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Development of bone marrow mesenchymal stem Cell culture in vitro

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Chinese Medical Journal 2012年第9期125卷 1650-1655页

作者： ZHANG Li PENG Li-pan WU Nan LI Le-ping Department of Gastrointestinal Surgery Provincial Hospital Affiliated to Shandong UniversityJinanShandong 250021China

Objective To review the in vitro development of bone marrow mesenchymal stem cells culture （BM-MSC）.Data sources The data cited in this review were mainly obtained from articles listed in Medline and PubMed. The search terms were “bone marrow mesenchymal stem cell” and “Cell culture”.Study selection Articles regarding the in vitro development of BM-MSCs culture, as well as the challenge of optimizing Cell culture environment in two-dimensional （2D） vs. 3D.Results Improving the culture conditions increases the proliferation and reduces the differentiation. Optimal values for many culture parameters remain to be identified. Expansion of BM-MSCs under defined conditions remains challenging, including the development of optimal culture conditions for BMSC and large-volume production systems.Conclusions Expansion of BM-MSCs under defined conditions remains challenges, including the development of optimal culture conditions for BMSC and scale-up to large-volume production systems. Optimal values for many culture parameters remain to be identified.

关键词： bone marrow mesenchymal stem cells Cell culture cell differentiation in vitro

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Biomimetic Graphene-Based 3D Scaffold for Long-Term Cell culture and Real-Time Electrochemical Monitoring

Biomimetic Graphene-Based 3D Scaffold for Long-Term Cell Cul...

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第十届全国化学生物学学术会议

作者： Xue-Bo Hu Yan-Ling Liu Hai-Wei Zhang Wei-Hua Huang Key Laboratory of Analytical Chemistry for Biology and Medicine （Ministry of Education） College of Chemistry and Molecular SciencesWuhan University

It is particularly challenging to construct a multifunctional 3D platform concurrently integrated with the function of Cell culture and electrochemical sensing.[1] Herein, we developed a 3-aminophenylboronic acid（APBA） functionalized graphene foam（GF） network, which combines the biomimetic property of APBA with the electrochemical properties of GF. The biomimetic network can serves as a 3D scaffold to culture cells for a long period with high viability（Figure 1a） and simultaneously as an electrode for highly sensitive electrochemical sensing of H2S（Figure 1b）. This allows monitoring of H2S released from the cells cultured on the 3D scaffold in real time. It is anticipated that this platform will open a new route for study towards cellular behaviors under 3D vivo-like microenvironments to reflect in vivo cell functions.

关键词： Graphene foam biomimetic scaffold Cell culture electrochemical monitoring

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Microfabrication of Bubbular Cavities in PDMS for cell Sorting and MicroCell culture Applications

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Journal of Bionic Engineering 2008年第4期5卷 308-316页

作者： Ut-Binh T. Giang Michael R. King Lisa A. DeLouise Department of Biomedical Engineering University of Rochester Department of Biomedical Engineering Cornell University Department of Dermatology University of Rochester Medical Center

We describe a novel technique, low surface energy Gas Expansion Molding （GEM）, to fabricate microbubble arrays in polydimethylsiloxane （PDMS） which are incorporated into parallel plate flow chambers and tested in cell sorting and microcell cuTture applications. This architecture confers several operational advantages that distinguish this technology approach from currently used methods. Herein we describe the GEM process and the parameters that are used to control microbubble formation and a Vacuum-Assisted Coating （VAC） process developed to selectively and spatially alter the PDMS surface chemistry in the wells and on the microchannel surface. We describe results from microflow image visualization studies conducted to investigate fluid streams above and within microbubble wells and conclude with a discussion of Cell culture studies in PDMS.

关键词： polydimethylsiloxane microfabrication Cell culture cell sorting molding

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An Innovative Design of Incubator Structure for Cell culture

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Medicinal Plant 2023年第3期14卷 105-107页

作者： Shanshan HE Zhongwei CHEN Ruonan HE Shiyi WU Qihuang LIN Engineering Research Center of Marine Biopharmaceutical Resource Fujian Province UniversityXiamen Medical CollegeXiamen 361023

In view of the problems of the traditional cell incubator,such as the small range of Cell culture types,the inability to adjust the internal space of the incubator according to needs,and the inconvenient sampling,this study innovatively designed a cell incubator structure.It proposed a new design concept that can solve the above-mentioned shortcomings.The cell incubator after the new structural modification can adjust the internal space structure of Cell culture by setting the bolt-fixed connection between the fixed plate and the vessel divider.It realizes the cultivation of various cells through refrigeration modules and heating modules.Through setting a sampling hole in the glass inner door,it is favorable for operators to take samples,making Cell culture more convenient and efficient.

关键词： cell incubator Innovative design Cell culture

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The Cell culture of forebrain neuron in zebrafish early stage embryos

The cell culture of forebrain neuron in zebrafish early stag...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Bin YANG,Gang PENG~* Institutes of Brain Science and State Key Laboratory of Medical Neuroscience,Fudan University,Shanghai 200032, China

＜正＞Objective The zebrafish（Danio rerio）,is an important model organism in nervous system development and animal behavior studies for its fast development and transparent embryos in early stage.Comparing to in vivo methods,in vitro Cell culture investigation approach has advantages in achieving higher resolution morphological observation and identifying typesof neurites with immunohistochemistry.Methods We use zebrafish early stage embryos to investigatefunctions of signal molecules and receptors for nervous development.We have developed methods involving bleaching,digestion,dissociation,plating of forebrain neurons and followed by immunohistochemistry.Results The Cell culture of forebrain neuron in zebrafish early stage embryos provides a new way of studying nervous development.

关键词： Danio rerio forebrain neuron Cell culture

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Binding of Orysata lectin induces an immune response in insect cells

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Insect Science 2022年第3期29卷 717-729页

作者： Pengyu Chen Kristof De Schutter Jarne Pauwels Kris Gevaert Els J.M.Van Damme Guy Smagghe Faculty of Bioscience Engineering Department of Plants and CropsGhent UniversityGhentBelgium Faculty of Bioscience Engineering Department of BiotechnologyGhent UniversityGhentBelgium Faculty of Medicine and Health Sciences Department of Biomolecular MedicineGhent UniversityGhentBelgium VIB Center for Medical Biotechnology GhentBelgium

In mammals,plant lectinshave been shown to possess immunomodulatory properties,acting in both the innate and adaptive immune system to modulate the production of mediators of the immune response,ultimately improving host defences.At present,knowledge of immunomodulatory effects of plant lectins in insects is scarce.Treatment of insect cells with the Orysa sativa lectin,Orysata,was previously reported to induce cell aggregation,mimicking the immune process of encapsulation.In this project we investigated the potential immunomodulatory effects of this mannose-binding lectin using Drosophila melanogaster S2 cells.Identification of the Orysata binding partners on the surface of S2 cells through a pull-down assay andproteomic analysis revealed 221 putative interactors,several of which were immunity-related proteins.Subsequent qPCR analysis revealed the upregulation of Toll-and immune deficiency(IMD)-regulated antimicrobial peptides(Drs,Mtk,AttA,and Dpt)and signal transducers(Rel and Hid)belonging to the IMD pathway.In addition,the iron-binding protein Transferrin 3 was identified as a putative interactor for Orysata,and treatment of S2 cells with Orysata was shown to reduce the intracellular iron concentration.All together,we believe these results offer a new perspective on the effects by which plant lectins influence insect cells and contribute to the study of their immunomodulatory properties.

关键词： Cell culture immune response iron uptake Orysata plant lectin pull-down

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PRIMARY Cell culture FOR EMBRYONIC CARDIAC MYOCYTES OF MEXICAN AXOLOTL AND DISTRIBUTION OF DESMIN AND VIMENTIN

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中国组织化学与细胞化学杂志 1999年第1期8卷 113-117,131页

作者： Zhu Yongze Robert W Zajdel Margaret E Fransen Larry F Lemanski Department of Anatomy Medical College of Yangzhou UniversityYangzhou225001 China Department of Anatomy and cell Biology State University of New YorkHealth Science Center atSyracuseSyracuseNY13210 U.S.A

Desmin and vimentin are major components of intermediate filament proteins in cardiac myocytes. We developed a primary Cell culture method for cardiac myocytes of axolotl embryos. Cardiac myocytes of embryonic stage 39 were cultured for 1-14 days. Myocytes showed spontaneous contractions (15-30 beats/min) after 48-72 hours in culture, round shape and large irregular projections. Desmin and vimentin were observed in the cultured myocytes by means of immunofluorescent staining in combination with immunofluorescent microscopy. Immunofluorescent staining of the cultured cardiac myocytes after different lengths of time in culture(3,6,9 days) showed that vimentin staining was stronger than desmin staining during the early stages of culture (3 days). The myocytes exhibited various forms of staining, including parallel lines and interconnected networks. Some lines showed regular striation; most of the myofibrils were arranged in parallel arrays along the cell's long axis. Both desmin and vimentin in the cell appeared to encirele the Z lines and to link myofibrils laterally at the Z lines.

关键词： Desmin Vimentin Cell culture Immunocytochemistry Axolotl

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