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Facile and rapid generation of micro-collagen-gel array for high throughput long-term single cell 3d culture

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Facile and rapid generation of micro-collagen-gel array for ...

Epithelial Cells in 2d and 3d cultures Exhibit Large differences in Higher-order Genomic Interactions

第八届全国化学生物学学术会议

作者： Shasha Jia Zhichao Guan Zhi Zhu Chaoyong James Yang The Key Laboratory for Chemical Biology of Fujian Province Key Laboratory of Analytical ScienceDepartment of Chemical BiologyCollege of Chemistry and Chemical EngineeringXiamen University

＜正＞Heterogeneity in cell populations po ses a major obstacle to understanding complex biological *** need for scalable analysis of single cell is particularly acute in the investigation of cell behavior and ***,Thr... 详细信息

关键词： single cell 3d culture high throughput

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Genomics, Proteomics & Bioinformatics 2022年第1期20卷 101-109页

作者： Xin Liu Qiu Sun Qi Wang Chuansheng Hu Xuecheng Chen Hua Li daniel M.Czajkowsky Zhifeng Shao State Key Laboratory for Oncogenes and Bio-Id Center School of Biomedical EngineeringShanghai Jiao Tong UniversityShanghai 200240China Shanghai Center for Systems Biomedicine Shanghai Jiao Tong UniversityShanghai 200240China Translational Medical Center for Stem Cell Therapy&Institute for Regenerative Medicine Shanghai East HospitalSchool of Life Science and TechnologyShanghai Key Laboratory of Signaling and Disease ResearchTongji UniversityShanghai 200092China

Recent studies have characterized the genomic structures of many eukaryotic cells,often focusing on their relation to gene ***,these studies have largely investigated cells grown in 2d cultures,although the transcriptomes of 3d-cultured cells are generally closer to their in vivo *** examine the effects of spatial constraints on chromosome conformation,we investigated the genomic architecture of mouse hepatocytes grown in 2d and 3d cultures using in situ *** results reveal significant differences in higher-order genomic interactions,notably in compartment identity and strength as well as in topologically associating domain(TAd)-TAd interactions,but only minor differences are found at the TAd *** RNA-seq analysis reveals up-regulated expression of genes involved in physiological hepatocyte functions in the 3d-cultured *** genes are associated with a subset of structural changes,suggesting that differences in genomic structure are critically important for transcriptional ***,there are also many structural differences that are not directly associated with changes in gene expression,whose cause remains to be ***,our results indicate that growth in 3d significantly alters higher-order genomic interactions,which may be consequential for a subset of genes that are important for the physiological functioning of the cell.

关键词： 3d culture In situ Hi-C Chromosome conformation Compartment TAd

Recent advances in the development of in vitro liver models for hepatotoxicity testing

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Bio-design and Manufacturing 2021年第4期4卷 717-734页

作者： Siming Lu Jingwei Zhang Sha Lin danfeng Zheng Yan Shen Jiale Qin Yangyang Li Shuqi Wang department of Laboratory Medicine The First Affiliated HospitalZhejiang University School of MedicineHangzhou 310003China State Key Laboratory for diagnosis and Treatment of Infectious diseases Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesThe First Affiliated HospitalNational Clinical Research Center for Infectious DiseasesZhejiang University School of MedicineHangzhou 310003China Institute of Translational Medicine Zhejiang UniversityHangzhou 310029China Key Laboratory of Women’s Reproductive Health of Zhejiang Province Women’s HospitalZhejiang University School of MedicineHangzhou 310006China Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province Zhejiang University School of MedicineHangzhou 310027China

Liver injury is a common cause of drug approval withdrawal during drug development,pre-clinical research,and clinical *** not properly treated,patients with severe liver injury can suffer from acute liver failure or even ***,utilization of the convenient in vitro hepatotoxicity assessment model for early detection of drug-induced hepatotoxicity is vital for drug development and safe personalized ***(e.g.,hydrogels,nanofibers,decellularized liver matrix)and bioengineering technologies(e.g.,microarrays,micropatterns,3d printing,and microfluidics)have been applied for in vitro hepatotoxicity assessment *** review summarizes the structure and functions of the liver as well as the components of in vitro hepatotoxicity assessment *** addition,it highlights the latest advances in developing hepatotoxicity models with the ultimate goal of further clinical translation.

关键词： Liver injury drug toxicity In vitro model 3d culture

Polyurethane scaffold-based 3d lung cancer model recapitulates in vivo tumor biological behavior for nanoparticulate drug screening

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Regenerative Biomaterials 2023年第1期10卷 1618-1631页

作者： Lu Sun Xiaofei Wang Yushui He Boran Chen Baoyin Shan Jinlong Yang Ruoran Wang Xihang Zeng Jiehua Li Hong Tan Ruichao Liang department of Targeting Therapy&Immunology Department of Radiation OncologyCancer CenterWest China HospitalSichuan UniversityChengdu 610041People's Republic of China department of Medical Polymer Materials Department of Artificial OrganismCollege of Polymer Science and EngineeringState Key Laboratory of Polymer Materials EngineeringSichuan UniversityChengdu 610065People’s Republic of China department of Neurosurgery West China HospitalSichuan UniversityChengdu 610041People’s Republic of China

Lung cancer is the leading cause of cancer mortality *** studies in lung cancer hold the promise of screening for effective antitumor agents,but mechanistic studies and drug discovery based on 2d cell models have a high failure rate in getting to the ***,there is an urgent need to explore more reliable and effective in vitro lung cancer ***,we prepared a series of three-dimensional(3d)waterborne biodegradable polyurethane(WBPU)scaffolds as substrates to establish biomimetic tumor models in *** 3d WBPU scaffolds were porous and could absorb large amounts of free water,facilitating the exchange of substances(nutrients and metabolic waste)and cell *** scaffolds at wet state could simulate the mechanics(elastic modulus∼1.9 kPa)and morphology(porous structures)of lung tissue and exhibit good biocompatibility.A549 lung cancer cells showed adherent growth pattern and rapidly formed 3d spheroids on WBPU *** results showed that the scaffold-based 3d lung cancer model promoted the expression of anti-apoptotic and epithelial-mesenchymal transition-related genes,giving it a more moderate growth and adhesion pattern compared to 2d *** addition,WBPU scaffold-established 3d lung cancer model revealed a closer expression of proteins to in vivo tumor,including tumor stem cell markers,cell proliferation,apoptosis,invasion and tumor resistance *** on these features,we further demonstrated that the 3d lung cancer model established by the WBPU scaffold was very similar to the in vivo tumor in terms of both resistance and tolerance to nanoparticulate *** together,WBPU scaffold-based lung cancer model could better mimic the growth,microenvironment and drug response of tumor in *** emerging 3d culture system holds promise to shorten the formulation cycle of individualized treatments and reduce the use of animals while providing valid research data for clinical trials.

关键词： WBPU scaffolds 3d culture lung cancer biomimetic nanoparticulate drug screening

Recent advances in chemically defined and tunable hydrogel platforms for organoid culture

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Bio-design and Manufacturing 2021年第3期4卷 641-674页

作者： Tarun Agarwal Nehar Celikkin Marco Costantini Tapas K.Maiti Pooyan Makvandi department of Biotechnology Indian Institute of Technology KharagpurKharagpurWest Bengal 721302India Institute of Physical Chemistry-Polish Academy of Sciences WarsawPoland Centre for Micro-BioRobotics Istituto Italiano di TecnologiaViale Rinaldo Piaggio 3456025 PontederaPisaItaly

Recent developments in organoid culture technologies have made it possible to closely recapitulate intrinsic characteristics of different tissues under in vitro *** organoids act as a translational bridge between the traditional 2d/3d cultures and the in vivo models for studying the tissue development processes,disease modeling,and drug *** and tissue-specific extracellular matrix have been shown to support organoid development,efficiently;however,their chemically undefined nature,non-tunable properties,and associated batch-to-batch variations often limit reproducibility of the assembly *** this regard,chemically defined platforms offer wider opportunities to optimize and recreate tissue-specific *** present review delineates the current research trends in this sphere,focusing on material perspective and the target tissues(e.g.,neural,liver,pancreatic,renal,and intestinal).The review winds up with a discussion on the current limitations and future perspective to provide a basis for future research.

关键词： Organoid 3d culture Biomaterials Chemically defined hydrogels Cell-instructive microenvironment

Long-term in-vitro culture and subculture of the hemocytes of swimming crab Portunus trituberculatus

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Journal of Oceanology and Limnology 2023年第5期41卷 1918-1939页

作者： Liwen GUO Yaqi ZHAO Huarong GUO Key Laboratory of Marine Genetics and Breeding Ministry of EducationCollege of Marine Life SciencesOcean University of ChinaQingdao 266003China Institute of Evolution&Marine Biodiversity Ocean University of ChinaQingdao 266003China

Crab cell line,especially continuous crab cell line,can provide us a useful tool for studies on the virology,immunology,and molecular biology of ***,no continuous crab cell line has been available due to the lacking of suitable medium and the occurrence of *** this study,long-term in vitro culture conditions for both two-(2d)and three-dimensions(3d)were successfully developed for the circulating hemocytes of swimming crab Portunus trituberculatus,designated as PTH *** 2d culture,a novel crab basic medium in osmolarity of 990–1100 mOsm/kg was optimized for the first time,which is different from Leibovitz's L-15 medium in mainly the components of amino acids,containing double strengths of the contents of free amino acid mixture in the crab *** an optimal crab growth medium was developed by supplementing 5%fetal bovine serum,50-g/L yeast extract powder,20-μg/L basic fibroblast growth factor and epidermal growth factor into the optimal crab basic medium,and found that it could support a long-term survival of PTH cells in a healthy monolayer up to 347 days and partially break through the mitosis-arrest of crab cells evidenced by the obvious increase of proliferating potential detected in the 10-d primarily cultured PTH *** 2d cultured PTH cells could be successfully sub-cultured for 11 times by physical flushing method and well cryopreserved in liquid *** 3d culture,using the same crab growth medium,the PTH cell aggregates could be easily formed and healthily maintained on the surface of solidified Matrigel or in the ultra-low-attachment plate with a survival rate of 50%–60%on day *** work largely improved the primary culture and subculture of crab cells and will facilitate the establishment of continuous crab cell line.

关键词： crab Portunus trituberculatus hemocyte long-term cell culture subculture 3d culture

decellularized extracellular matrix mediates tissue construction and regeneration

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Frontiers of Medicine 2022年第1期16卷 56-82页

作者： Chuanqi Liu Ming Pei Qingfeng Li Yuanyuan Zhang department of Plastic and Burn Surgery West China HospitalSichuan UniversityChengdu 610041China department of Plastic and Reconstructive Surgery Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghai 200011China Stem Cell and Tissue Engineering Laboratory Department of OrthopaedicsWest Virginia UniversityMorgantownWV 26506USA Wake Forest Institute for Regenerative Medicine Wake Forest University Health SciencesWinston-SalemNC 27109USA

Contributing to organ formation and tissue regeneration,extracellular matrix(ECM)constituents provide tissue with three-dimensional(3d)structural integrity and cellular-function *** the crucial traits of the cellular microenvironment,ECM substitutes mediate cell–matrix interactions to prompt stem-cell proliferation and differentiation for 3d organoid construction in vitro or tissue regeneration in ***,these ECMs are often applied generically and have yet to be extensively developed for specific cell types in 3d *** cells also produce rich ECM,particularly stromal *** ECM improves 3d culture development in vitro and tissue remodeling during wound healing after implantation into the host as *** better insight into ECM derived from either tissue or cells that regulate 3d tissue reconstruction or organ regeneration helps us to select,produce,and implant the most suitable ECM and thus promote 3d organoid culture and tissue remodeling for in vivo ***,the decellularization methodologies and tissue/cell-derived ECM as scaffolds or cellular-growth supplements used in cell propagation and differentiation for 3d tissue culture in vitro are ***,current preclinical applications by which ECM components modulate the wound-healing process are reviewed.

关键词： decellularized extracellular matrix 3d culture organoids tissue repair

Integrin binding peptides facilitate growth and interconnected vascular-like network formation of rat primary cortical vascular endothelial cells in vitro

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Neural Regeneration Research 2023年第5期18卷 1052-1056页

作者： Ram Kuwar Xuejun Wen Ning Zhang dong Sun department of Anatomy and Neurobiology Medical College of Virginia CampusVirginia Commonwealth UniversityRichmondVAUSA department of Chemical and Life Science Engineering College of EngineeringVirginia Commonwealth UniversityRichmondVAUSA department of Biomedical Engineering College of EngineeringVirginia Commonwealth UniversityRichmondVAUSA

Neovascularization and angiogenesis in the brain are important physiological processes for normal brain development and repair/regeneration following insults. Integrins are cell surface adhesion receptors mediating important function of cells such as survival, growth and development during tissue organization, differentiation and organogenesis. In this study, we used an integrin-binding array platform to identify the important types of integrins and their binding peptides that facilitate adhesion, growth, development, and vascular-like network formation of rat primary brain microvascular endothelial cells. Brain microvascular endothelial cells were isolated from rat brain on post-natal day 7. Cells were cultured in a custom-designed integrin array system containing short synthetic peptides binding to 16 types of integrins commonly expressed on cells in vertebrates. After 7 days of culture, the brain microvascular endothelial cells were processed for immunostaining with markers for endothelial cells including von Willibrand factor and platelet endothelial cell adhesion molecule. 5-Bromo-2′-dexoyuridine was added to the culture at 48 hours prior to fixation to assess cell proliferation. Among 16 integrins tested, we found that α5β1, αvβ5 and αvβ8 greatly promoted proliferation of endothelial cells in culture. To investigate the effect of integrin-binding peptides in promoting neovascularization and angiogenesis, the binding peptides to the above three types of integrins were immobilized to our custom-designed hydrogel in three-dimensional(3 d) culture of brain microvascular endothelial cells with the addition of vascular endothelial growth factor. Following a 7-day 3 d culture, the culture was fixed and processed for double labeling of phalloidin with von Willibrand factor or platelet endothelial cell adhesion molecule and assessed under confocal microscopy. In the 3 d culture in hydrogels conjugated with the integrin-binding peptide, brain microvascular endothelial c

关键词： 3d culture angiogenesis brain microvascular endothelial cells hydrogel integrins platelet endothelial cell adhesion molecule(PECAM-1) vascular endothelial growth factor(VEGF) vascularization

Organoid technology for tissue engineering

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Journal of Molecular Cell Biology 2020年第8期12卷 569-579页

作者： Juan He Xiaoyu Zhang Xinyi Xia Ming Han Fei Li Chunfeng Li Yunguang Li dong Gao State Key Laboratory of Cell Biology Shanghai Key Laboratory of Molecular AndrologyCAS Center for Excellence in Molecular Cell ScienceShanghai Institute of Biochemistry and Cell BiologyChinese Academy of SciencesUniversity of Chinese Academy of SciencesShanghai 200031China Institute for Stem Cell and Regeneration Chinese Academy of SciencesBeijing 100101China

For centuries,attempts have been continuously made to artificially reconstitute counterparts of in vivo organs from their tissues or *** in the recent decade has organoid technology as a whole technological field systematically emerged and been shown to play important roles in tissue *** on their self-organizing capacities,stem cells of versatile organs,both harvested and induced,can form 3d structures that are structurally and functionally similar to their in vivo *** organoid models provide a powerful platform for elucidating the development mechanisms,modeling diseases,and screening drug *** this review,we will summarize the advances of this technology for generating various organoids of tissues from the three germ layers and discuss their drawbacks and prospects for tissue engineering.

关键词： organoid tissue engineering 3d culture