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Frontiers of Materials Science 2024年第2期18卷 129-137页

作者： Huaying Hao Lihong Sun Jiaxuan Chen Jun Liang State Key Laboratory of Food Nutrition and Safety Tianjin University of Science&TechnologyTianjin 300457China College of Light Industry Science and Engineering Tianjin University of Science&TechnologyTianjin 300457China College of Food Science and Engineering Tianjin University of Science&TechnologyTianjin 300457China

The in vitro expansion of stem cells is important for their application in different life science fields such as cellular tissue and organ *** objective of this paper was to achieve static cell culture in vitro through peptide hydrogel-supported microspheres(MSs).The peptides,with their gel-forming properties,microstructures,and mechanical strengths characterized,were found to have good support for the MSs and to be *** internal structures of poly(L-lactic acid)microspheres(PLLA-MSs)and polystyrene microspheres(PS-MSs)made in thelaboratory were observed and statistically analyzed in terms of particle size and pore size,following which the co-cultured MSs with cells were found to have good cell *** addition,three-dimensional(3D)culturing of cells was performed on the peptide and microcarrier composite scaffolds to measure cell viability and cell *** results showed that the peptides could be stimulated by the culture medium to self-assembly form a 3D fiber network *** the peptide-Ms composite scaffold-based cell culture system,further enhancement of the cell culture effect was *** peptide-Ms composite scaffolds have great potential for the application in 3D cell culture and in vitro cellexpansion.

关键词： microcarrier microsphere peptide hydrogel cell scaffold three-dimensional culture cell expansion

Chinese medicinal compound delisheng has satisfactory anti-tumor activity,and is associated with up-regulation of endostatin in human hepatocellular carcinoma cell line HepG2 in three-dimensional culture

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World Journal of Gastroenterology 2007年第41期13卷 5432-5439页

作者： Jie Cui Ke-Jun Nan Tao Tian Ya-Huan Guo Na Zhao Lin Wang Department of Medical Oncology The First Affiliated Hospital of The School of Medicine of Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China

AIM： TO investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng, 5-fluorouracil and adriamycin, and the possible molecular mechanisms of delisheng. METHODS： Human hepatocellular carcinoma HepG2 cells were cultured with a liquid overlay technique. After the formation of multicellular spheroids, morphology was analyzed by phase contrast microscopy, scanning electron microscopy and transmission electron microscopy. Sensitivity of HepG2 cells to delisheng, 5-fluorouracil and adriamycin was investigated by Ml-I- assay in multicelluar spheroids and monolayers. Vascular endothelial growth factor （VEGF） and endostatin expression were analyzed in multicellular spheroids treated with delisheng, 5-fluorouracil, adriamycin and negative control PBS, with immunohistochemical staining. RESULTS： Multicellular spheroids exhibited structural characteristics somewhat different to those in monolayers. The cells in three-dimensional cell culture turned out to be less sensitive to delisheng, 5-fluorouracil and adriamycin than the cells cultured in monolayer. This showed that delisheng had a satisfactory cells inhibition ratio compared to 5-fluorouracil and adriamycin. Immunohistochemical staining showed that VEGF and endostatin expression was positive during growth as multicellular spheroids, and endostatin expression in spheroids with treatment of delisheng was higher than that with 5-fluorouracil, adriamycin and PBS （139.35 ± 7.83, 159.23 ± 10.34, 162.83 ± 3.47 and 148.48 ± 11.06, P 〈 0.05）.CONCLUSION： Chinese medicine compound delisheng has satisfactory anti-tumor activity in HepG2 cells in three-dimensional culture, and the effects are associated with up-regulation of endostatin.

关键词： Delisheng Ginseng three-dimensional culture Multicellular resistance Endostatin

Isolation and Identification of Cancer Stem Cells from Human Osteosarcom by Serum-free three-dimensional culture Combined with Anticancer Drugs

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2010年第1期30卷 81-84页

作者：周松李锋肖骏熊伟方忠陈文坚牛鹏彦 Department of Orthopedics Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology

The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and *** primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate ***-cell alginate gel was cultured with serum-free DMEM/F12 ***(0.8μg/mL)was added to the medium to enrich *** cultured conventionally for 7 to 10 days,most of cells suspended in ...

关键词： three-dimensional culture serum-free culture epirubicin osteosarcoma cancer stem cells

Assessment of pancreatic carcinoma cell chemosensitivity using a three-dimensional culture system

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Chinese Medical Journal 2010年第14期123卷 1871-1877页

作者： LIAO Quan HU Ya ZHAO Yu-pei ZHOU Tao ZHANG Qiang Department of General Surgery Peking Union Medical College Hospital National Laboratory of Medical Molecular Biology Chinese Academy of Medical Sciences Beijing 100730 China

Background Monolayer cell culture models are the traditional culture models used for in vitro research of pancreatic carcinoma chemosensitivity. However, these models neglect the interactions between tumor cells and the impact of the tumor microenvironment. Such tumor cell monolayers poorly mimic the solid tumor microenvironment. The present study aimed to investigate the chemosensitivity characteristics of pancreatic cancer cells in a three-dimensional culture system by analyzing the differences in drug sensitivity between a scattered cell culture model and a multicellular spheroid culture model. Methods three pancreatic cancer cell lines （SW1990, ASPC-1 and PCT-3） were cultured in three-dimensional collagen gels as well as in traditional two-dimensional monolayers. The chemosensitivities of the pancreatic carcinoma cells to 5-fluorouracil （5-FU）, gemcitabine, and oxaliplatin in vitro were detected by both the Cell Counting Kit-8 test and the collagen gel droplet-embedded culture drug-sensitivity test. Results In the two-dimensional culture model, differences in the chemosensitivities of the cloned pancreatic carcinoma cells and scattered cells existed for some concentrations of 5-FU, gemcitabine and oxaliplatin. In the three-dimensional culture model, there were significant differences in the chemosensitivities of the pancreatic cancer cells between the scattered cells and multicellular spheroids （P 〈0.05）. Conclusion Pancreatic carcinoma cells exhibit multicellular resistance in three-dimensional cultures.

关键词： pancreatic carcinoma cell chemosensitivity three-dimensional culture

Modulation of the Nogo signaling pathway to overcome amyloid-β-mediated neurite inhibition in human pluripotent stem cell-derived neurites

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Neural Regeneration Research 2025年第9期20卷 2645-2654页

作者： Kirsty Goncalves Stefan Przyborski Department of Biosciences Durham UniversityDurhamUK Reprocell Europe Ltd. GlasgowUK

Neuronal cell death and the loss of connectivity are two of the primary pathological mechanisms underlying Alzheimer's *** accumulation of amyloid-βpeptides,a key hallmark of Alzheimer's disease,is believed to induce neuritic abnormalities,including reduced growth,extension,and abnormal growth cone morphology,all of which contribute to decreased ***,the precise cellular and molecular mechanisms governing this response remain *** this study,we used an innovative approach to demonstrate the effect of amyloid-βon neurite dynamics in both two-dimensional and three-dimensional cultu re systems,in order to provide more physiologically relevant culture *** utilized various methodologies,including the addition of exogenous amyloid-βpeptides to the culture medium,growth substrate coating,and the utilization of human-induced pluripotent stem cell technology,to investigate the effect of endogenous amyloid-βsecretion on neurite outgrowth,thus paving the way for potential future applications in personalized ***,we also explore the involvement of the Nogo signaling cascade in amyloid-β-induced neurite *** demonstrate that inhibition of downstream ROCK and RhoA components of the Nogo signaling pathway,achieved through modulation with Y-27632(a ROCK inhibitor)and Ibuprofen(a Rho A inhibitor),respectively,can restore and even enhance neuronal connectivity in the presence of amyloid-β.In summary,this study not only presents a novel culture approach that offers insights into the biological process of neurite growth and inhibition,but also proposes a specific mechanism for reduced neural connectivity in the presence of amyloid-βpeptides,along with potential intervention points to restore neurite ***,we aim to establish a culture system that has the potential to serve as an assay for measuring preclinical,predictive outcomes of drugs and their ability to promote neurite outgrowth,both generally and in a patie

关键词： Alzheimer's disease induced pluripotent stem cell neurite outgrowth neuron Nogo Rho A ROCK stem cell three-dimensional culture

three-dimensional cultures of human endometrial cells on Matrigel mimic in vivo morphology

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Chinese Medical Journal 2012年第5期125卷 863-868页

作者： ZHU Hai-yan WANG Jun-xia TONG Xiao-mei XU Wei-hai JIANG Ling-ying JING Xiao-ying YANG Ling-yun ZHOU Feng ZHANG Song-ying Assisted Reproduction Unit Department of Obstetrics and GynecologySir Run Run Shaw HospitalCollege of MedicineZhejiang UniversityHangzhouZhejiang 310016China

Background The regulation of endometrial physiology and morphogenesis by the paracrine effectors has been well established using in vivo studies. A more complete understanding of the endometrial function has been delayed due, in part, to a lack of appropriate culture models. In this study, we aimed to simulate the in vivo three-dimensional （3-D） growth pattern of endometrial cells using a 3-D in vitro culture system.

关键词： Matrigel endometrium three-dimensional culture morphology

Engineered three-dimensional rabbit oral epithelial–mesenchymal–muscular hybrid sheets

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International Journal of Oral Science 2016年第3期8卷 145-154页

作者： Shigeki Yamane Kazunari Higa Takashi Umezawa Masamitsu Serikawa Jun Shimazaki Shinichi Abe Department of Anatomy Tokyo Dental College Tokyo Japan Department of Ophthalmology/Cornea Center Tokyo Dental College Chiba Japan

Regenerative muscles are required for swallowing and mastication, and are important for functional recovery from diseases involving oral muscular defects. Therefore, we generated three-layer hybrid sheets, similar to oral mucosal structures containing submucosal muscles, using rabbit oral mucosa epithelial, mesenchymal, and myoblastic progenitor cells, and examined the structural proteins. Each cell type was obtained from rabbit oral mucosa using enzymatic digestion. Isolated mesenchymal and myoblastic cells were multi-differentiated into osteoblasts, adipocytes, and chondrocytes or myotubes. Isolated epithelial cells were cultured on collagen gels containing isolated mesenchymal cells for 2 weeks, and these epithelial-mesenchymal cell sheets were laminated onto myoblastic cell sheets. The engineered hybrid sheets were multi-stratified in the epithelial and myoblastic layers in a time-dependent manner, expressing intermediate cytoskeletal filament proteins of epithelium and muscle. Hybrid sheets also expressed extracellular matrix basement membrane proteins. Immature cell markers for epithelial and myoblastic cells were observed continuously in hybrid sheet cultures. We established engineered three-dimensional rabbit oral mucosa hybrid sheets containing each immature cell type in vitro.

关键词： mesenchymal stem cell multi-differentiation myoblast oral mucosa three-dimensional culture

Efficient expansion of rare human circulating hematopoietic stem/progenitor cells in steady-state blood using a polypeptide-forming 3D culture

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Protein & Cell 2022年第11期13卷 808-824页

作者： Yulin Xu Xiangjun Zeng Mingming Zhang Binsheng Wang Xin Guo Wei Shan Shuyang Cai Qian Luo Honghu Li Xia Li Xue Li Hao Zhang Limengmeng Wang Yu Lin Lizhen Liu Yanwei Li Meng Zhang Xiaohong Yu Pengxu Qian He Huang Bone Marrow Transplantation Center First Affiliated HospitalZhejiang University School of MedicineHangzhou 310012China Liangzhu Laboratory Zhejiang University Medical CenterHangzhou 311121China Institute of Hematology Zhejiang UniversityHangzhou 310012China Zhejiang Province Engineering Laboratory for Stem Cell and Immunity Therapy Hangzhou 310012China Core Facilities Zhejiang University School of MedicineHangzhou 310012China Dr.Li Dak Sum&Yip Yio Chin Center for Stem Cell and Regenerative Medicine Zhejiang UniversityHangzhou 310012China

Although widely applied in treating hematopoietic malignancies,transplantation of hematopoietic stem/progenitor cells(HSPCs)is impeded by HSPC *** circulating HSPCs(cHSPCs)in steady-state blood could be used as an alternative source remains largely *** we develop a three-dimensional culture system(3DCS)including arginine,glycine,aspartate,and a series of ***-day culture of peripheral blood mononuclear cells(PBMNCs)in 3DCS led to 125-and 70-fold increase of the frequency and number of CD34+***,3DCS-expanded cHSPCs exhibited the similar reconstitution rate com-pared to CD34+HSPCs in bone ***,3DCS fabricated an immunomodulatory niche,secreting cytokines as TNF to support cHSPC survival and ***,3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC *** 3DCS successfully expands rare cHSPCs,providing an alternative source for the HSPC therapy,particularly for the patients/donors who have failed in HSPC mobilization.

关键词： hematopoietic stem/progenitor cell transplantation peripheral blood mononuclear cells expansion mobilization three-dimensional culture selfrenewal and multilineage differentiation

The three-dimensional Collagen Scaffold Improves the Sternness of Rat Bone Marrow Mesenchymal Stem Cells

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Journal of Genetics and Genomics 2012年第12期39卷 633-641页

作者： Sufang Han Yannan Zhao Zhifeng Xiao Jin Han Bing Chen Lei Chen Jianwu Dai State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of Sciences University of Chinese Academy of Sciences

Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） substrate in vitro. There are indications that they may simultaneously lose their sternness and multipotentiality as the result of prolonged 2D culture. In this study, we used three-dimensional （3D） collagen scaffolds as rat MSCs cartier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs. The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation. More importantly, compared to MSCs under 2D culture, 3D MSCs significantly maintained higher expression levels of stemness genes （Oct4, Sox2, Rex-1 and Nanog）, yielded high frequencies of colony-forming units-fibroblastic （CFU-F） and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction. Thus, 3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.

关键词： three-dimensional culture Mesenchymal stem ceils Collagen scaffold Stemness

Morphological properties and proliferation analysis of olfactory ensheathing cells seeded onto three-dimensional collagen-heparan sulfate biological scaffolds

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Neural Regeneration Research 2012年第16期7卷 1213-1219页

作者： Na Liu Zhouping Tang Zhiyuan Yu Minjie Xie Yu Zhang Erfang Yang Shabei Xu Department of Neurology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 HubeiProvince China

This study aimed to examine the differences in the morphological properties and proliferation of olfactory ensheathing cells in three-dimensional culture on collagen-heparan sulfate biological scaffolds and in two-dimensional culture on common flat culture plates. The proliferation rate of olfactory ensheathing cells in three-dimensional culture was higher than that in two-dimensional culture, as detected by an M-I-r assay. In addition, more than half of the olfactory ensheathing cells subcultured using the trypsinization method in three-dimensional culture displayed a spindly Schwann cell-like morphology with extremely long processes, while they showed a flat astrocyte-like morphology in two-dimensional culture. Moreover, spindle-shaped olfactory ensheathing cells tended to adopt an elongated bipolar morphology under both culture conditions. Experimental findings indicate that the morphological properties and proliferation of olfactory ensheathing cells in three-dimensional culture on collagen-heparan sulfate biological scaffolds are better than those in two-dimensional culture.

关键词： morphological properties cell proliferation biological scaffold olfactory ensheathing cells three-dimensional culture neural regeneration