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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

Constructing thermoresponsive PNIPAM-based microcarriers for cell culture and enzyme-free cell harvesting

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Chinese Chemical Letters 2024年第7期35卷 412-416页

作者： Yunan Yuan Zhimin Luo Jie Chen Chaoliang He Kai Hao Huayu Tian CAS Key Laboratory of Polymer Ecomaterials Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchun 130022China School of Applied Chemistry and Engineering University of Science and Technology of ChinaHefei 230026China College of Chemistry and Chemical Engineering Xiamen UniversityXiamen 361005China Jilin Biomedical Polymers Engineering Laboratory Changchun 130022China Innovation Laboratory for Sciences and Technologies of Energy Materials of Fujian Province(IKKEM) Xiamen 361005China

The development of large-scale cell cultivation and non-invasive cell harvesting is highly desired in various fields,including biological regeneration and pharmaceutical *** using traditional microcarriers for cell culture,trypsinization is often necessary during cell collection,leading to partial cells *** this work,we developed a thermoresponsive glass microcarrier modified with poly(γ-propargyl-L-glutamate)(PPLG)and poly(N-isopropylacrylamide)(PNIPAM).We utilized these microcarriers for three-dimensional cell culture and enzyme-free cell harvesting,and the results indicated that the prepared microcarriers exhibited excellent non-invasive cell culture performance.

关键词： Microcarrier Temperature-responese PNIPAM Polymers cell culture

Microfluidic generation of Buddha beads-like microcarriers for cell culture

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Science China Materials 2017年第9期60卷 857-865页

作者：王洁邹旻含孙灵钰程瑶商珞然付繁繁赵远锦 State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China

The fabrication of functional microcarriers capable of achieving in vivo-like three-dimensional cell culture is important for many tissue engineering applications. Here,inspired by the structure of Buddha beads, which are generally composed of moveable beads strung on a rope, we present novel cell microcarriers with controllable macropores and heterogeneous microstructures by using a capillary array microfluidic technology. Microfibers with a string of moveable and releasable microcarriers could be achieved by an immediate gelation reaction of sodium alginate spinning and subsequent polymerization of cell-dispersed gelatin methacrylate emulsification. The sizes of the microcarriers and their inner macropores could be well tailored by adjusting the flow rates of the microfluidic phases; this was of great importance in guaranteeing a sufficient supply of nutrients during cell culture. In addition, by infusing multiple cell-dispersed pregel solutions into the capillaries, the microcarriers with spatially heterogeneous cell encapsulations for mimicking physiological structures and functions could also be achieved.

关键词： microfluidics microcarrier cell culture microfiber emulsion

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

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 *** study aimed to investigate the effects of SP on growth of fibroblast-like cells derived from rabbit bile *** Fibroblast-like cells derived from rabbit bile duct were identified and divided randomly into control and experimental ***-treated cells at different concentrations of 10^-9-10^-5 mol/L and control group were incubated,respectively,for 48 *** incubating,the effects of SP on cell proliferation were assessed by cell counts and MTT *** rate （AR） of cells was measured by flow *** cultured rabbit bile duct cells were fibroblast-like in morphology,and these cells were stained positively for vimentin and negatively for *** 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/*** 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 *** under transmission electron microscopy revealed rough endoplasmic reticulum and prominent Golgi complexes after SP *** 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

Complete Genomic Sequence of Transmissible Gastroenteritis Virus TS and 3' End Sequence Characterization Following cell culture

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Virologica Sinica 2010年第3期25卷 213-224页

作者： Jian-qiang LI Jie CHENG Xi LAN Xue-rui LI Wei LI Xiang-ping YIN Bao-yu LI Bin YANG Zhi-yong LI Yun ZHANG Ji-xing LIU Key laboratory of Animal Virology of Ministry of Agriculture State Key Laboratory of Veterinary Etiological Biology Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences Lanzhou 730046 China Lanzou Institute of Biological Products Lanzhou 730070 China

The complete genome sequence of transmissible Gastroenteritis virus (TGEV) strain TS, previously isolated from Gansu province, was cloned and compared with published sequence data from other TGEV strains. Phylogenetic tree analysis based on the amino acid and nucleotide sequences of the S gene showed that the TGEV strains were divided into 3 clusters. TGEV TS showed a close evolutionary relationship to the American Miller cluster but had a 5' non-translated region (NTR) sequence closely related to the American Purdue cluster. Continued culture in different cell types indicated that TGEV TS virulence could be attenuated after fifty passages in Porcine kidney (PK-15) cells, and that the Porcine kidney cell line IB-RS-2 (IBRS) was not suitable for culture of the TGEV strain TS.

关键词： Cloning Complete genome Gastroenteritis virus (TGEV) cell culture

Increased production of recombinant prourokinase with porous microcarrier cell culture by periodic pressure oscillation in a stirred tank reactor

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High Technology Letters 2006年第3期12卷 311-317页

作者：胡显文 Gao Lihua Li Zuohu Xiao Chengzu Xu Zhaoping Department of cell Engineering Institute of Bioteehnology Academy of Military Medical Sciences Beijing 100071 P.R. China State Key Laboratory of Biochemical Engineering Institute oi Process Engineering Chinese Academy of Sciences Beijing 100080 P. R. China

An rCHO cell line expressing recombinant human prourokinase （pro-UK） at the level of 5μg/ 10^6cells/d was cultivated on Cytopore cellulose porous microcarriers in a 7.5L Biostat CT stirred tank reactor. A periodic pressure oscillation of 0.04 MPa and 0.04 Hz was adopted to introduce a physical stimulus on the rCHO cells and to improve mass transfer characteristic between cells and medium in the process of porous microcarrier CHO cell culture. Compared to constant pressure culture, the oscillation culture didn＇t influence specific cell growth rate significantly, but could enhance the pro-UK specific production by 10% - 40%, and reduce production of lactate by 10% - 30%. In the perfusion culture of recombinant CHO cell with serum-free medium for 67 days, cell density could reach 2.64×10^7/ml, the maximal prourokinase concentration in harvested supernatant was about 118mg/L, a total of 21.1 grams of prourokinase was produced in 313 liters of supernatant. In conclusion, the perfusion cell culture with periodic pressure oscillation can enhance the production of recombinant protein and increase the reactor specific productivity.

关键词： cell culture porous microcarrier prourokinase periodic pressure oscillation

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 *** proposed a new design concept that can solve the above-mentioned *** 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 *** realizes the cultivation of various cells through refrigeration modules and heating *** 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

THE RESEARCH OF METHODS FOR SF9 cell culture TECHNOLOGY

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泰山医学院学报 1999年第1期 37-39页

作者： Zhu Changjun 1,Zhao Jian 2,Zhang Lining 3,Qu Xun 1,Liu Guiqiu 1,Song Wengang 1 (1 Taishan Medical College,271000 2 Affiliated Hospital of Shandong Medical University 3 Shandong Medical University)

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关键词： SF9 cell cell culture