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Differential gene expression during capillary morphogenesis in a microcarrier-based three-dimensional in vitro model of angiogenesis with focus on chemokines and chemokine receptors

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World Journal of Gastroenterology 2005年第15期11卷 2283-2290页

作者： Xi-TaiSun Min-YueZhang ChangShu QiangLi Xiao-GuiYan NiCheng Yu-DongQiu Yi-TaoDing DepartmentofHepatobiliarySurgery AffiliatedDrumTowerHospitalofMedicalCollegeHepatobiliaryResearchInstituteNanjingUniversityNanjing210008JiangsuProvinceChina StateKeyLaboratoryofPharmaceuticalBiotechnology DepartmentofBiochemistryNanjingUniversityNanjing210009JiangsuProvinceChina StateKeyLaboratoryofPharmaceuticalBiotechnology ModelAnimalResearchCenterDepartmentofBiochemistryNanjingUniversityNanjing210009JiangsuProvinceChina

AIM:To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogeness system with affymetrix oligonucleotide array. METHODS: A microcarrier-based in vitro angiogenesis system was developed, in which ECs migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed networks. The total RNA samples from the HMVECs at the selected time points (0.5, 24, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the softwares provided by the manufacturers. The expression patterns of some genes were validated and confirmed by semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines and chemokine receptors was specially examined and their functional implications were analyzed. RESULTS: A total of 1 961 genes were up- or downreg-ulated two-folds or above, and among them, 468 genes were up- or down-regulated three-folds or above. The regulated genes could be grouped into categories based on their molecular functions, and were also clustered into six groups based on their patterns of expression. As for chemokines and chemokine receptors, CXCL1/GRO-α, CXCL2/GRO-β, CXCL5/ENA-78, CXCL6/GCP2, IL-8/CXCL8, CXCL12/SDF-1, CXCL9/Mig, CXC11/ITAC, OOCL1/fractalkine, CCL2/MCP-1, CCL3, CCL5/RANTES, CCL7, CCL15, CCL21, CCL23, CCL28, and CCR1, CCR9, CXCR4 were identified. Moreover, these genes demonstrated different changing patterns during the capillary morphogenesis, which implied that they might have different roles in the sequential process. Among the chemokines identified, CCL2/MCP-1, CCL5/RANTES and CX3CL1 were specially up-regulated at the 24-h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early s

关键词： Angiogenesis in vitro model Endothelial cell Oligonucleotide array

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Validation of an in vitro model to Study Human Cartilage Responses to Compression

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Engineering（科研） 2012年第10期4卷 61-64页

作者： Natalia de Isla Céline Huselstein Didier Mainard Jean-Francois Stoltz CNRS UMR 7561 BiopoleFaculte de MédecineUniversite de LorraineVandoeuvreFrance UTCT CHU de NancyVandoeuvreFrance COT CHU de NancyVandoeuvreFrance

The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee were compressed at 1MPa/1Hz for 7 hours (30 min ON, 30 min OFF) under normoxia (5% CO2, 21% O2) or hypoxia (5% CO2, 5% O2). Cell viability was analyzed while nitric oxide (NO) and glycosaminoglycans (GAG) release was assayed in culture media. Mechanical stimulation increased NO production and GAG release by human cartilage explants under normoxia and hypoxia culture. In normoxia and hypoxia conditions, mechanical stimulation alters human OA cartilage metabolism. There is also, an increase in matrix degradation after compression, as shown by levels of GAG found in culture media. This study put in evidence the importance of mechanical compression in the progression of the osteoarthritis and present and in vitro model for mechanobiological and pharmacological studies.

关键词： Cartilage Compression in vitro model Nitric Oxide GAG

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Evaluating effects of gypenosides and soyasaponins on differentiation of neural stem cells as an in vitro model

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Journal of Shanghai University(English Edition) 2008年第1期12卷 91-94页

作者：吴婕文铁桥 Laboratory of Neural Molecular Biology School of Life Sciences Shanghai University Shanghai 200444 P. R. China

Neural stem cell has a potential to differentiate into neurons, astrocytes and oligodendrocytes. It provides an in vitro model to screen herbal medicines on the cellular differentiation and development level. In this work, active component from gypenosides and soyasaponins was prepared to investigate their effects on the differentiation of neural stem cells.. Both gypenosides and soyasaponins promote the differentiation of neural stem cells. This method provides speed and practicality for screening effective herbal medicine. It is well suited for studying the mechanism of cell differentiation and development.

关键词： neural stem cell gypenosides soyasaponins herbal medicine in vitro model

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An In vitro Biological model for Evaluation of the Anti-Dandruff Performance of Hair Care Products

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China Detergent & Cosmetics 2018年第1期3卷 69-73页

作者： Chen Mo Jiang Dan Huang Liang Zhang Liping Guangzhou Liby Enterprise Group co. Ltd.

An in vitro biological model for evaluation of the anti-dandruff performance of hair care products was established. In this model, Malasseziafurfur(***) was used as indicator bacteria with pigskin as *** model functions in following steps: pretreatment for the substrate; inoculation of indicator bacteria;measuring the bacteriostatic efficacy by the contact plates after washing the pigskin with the sample shampoo;calculation of the effective index P; evaluation of the anti-dandruff performance by the effective index P. Three kinds of anti-dandruff agents including BPT, ZPT and OCT were tested by the model. Results showed that the order for antibacterial efficacy of the anti-dandruff agents within the mass fraction range of 0.1 %~2.0% was BPT>ZPT>OCT. Results of the evaluation on each type of shampoo showed that: one displays no antidandruff efficacy; three displays medium anti-dandruff efficacy; two displays strong anti-dandruff efficacy.

关键词： anti-dandruff shampoo Malassezia furfur in vitro model

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Normal mouse intestinal epithelium as a model in vitro for invasion by Trichinella spiralis infective larvae

Normal mouse intestinal epithelium as a model in vitro for i...

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中国动物学会寄生虫学专业委员会第十三次全国学术会议暨第四次国际寄生虫学学术研讨会

作者： Hui-Jun Ren Jing Cui Zhong-Quan Wang Department of Parasitology Medical College Zhengzhou University

It has been known for many years that Trichinella spiralis initiates infection by penetrating the columnar epithelium of the small intestine;however,the mechanisms used by the parasite in the establishment of its intramulticellular niche in the intestine are *** the previous observations indicated that invasion also occurs in vitro when the infective larvae are inoculated onto cultures of intestinal epithelial cells(e.g.,human colonic carcinoma cell line Caco-2,HCT-8),a normal readily manipulated in vitro model has not been established because of difficulties in the culture of primary intestinal epithelial cells(IECs).In this study,we described a normal intestinal epithelium model in which the *** infective larvae were shown to invade the monolayers of normal mouse IECs in *** penetrated cells and migrated through them,leaving trails of dead *** cells derived from intestinal crypts of fetal mouse small intestine had the ability to proliferate continuously and express specific cytokeratins as well as intestinal functional cell ***,the normal mouse IECs were susceptible to invasion of ***,and those cells invaded by this parasite were heavily loaded with *** larval excretory-secretory antigens which were recognized by mouse immune *** normal intestinal epithelial model of invasion mimicking the natural environment in vivo provides a readily manipulated and controlled system to further investigate the process as well as the mechanisms by which *** establishes its intestinal niche.

关键词： Trichinella spiralis in vitro model intestinal epithelial cells Invasion mice

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Effects of natural-cerebrolysin-containing serum on neurotoxicity and synaptogenesis in amyloid-beta 1-40-induced Alzheimer's disease in vitro models

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Neural Regeneration Research 2009年第2期4卷 85-91页

作者： Yinghong Li Zhengzhi Wu Andrew C. J. HuangO Ming Li XiaoLi Zhang Jiguo Wang Shenzhen Hospital Southern Medical University Shenzhen 518033 Guangdong Province China Larry Hillblom Islet Research Center David Geffen School of Medicine UCLA Los Angeles CA 90024 USA

BACKGROUND： Neuronal loss, synapse mutilation, and increasing malnourished axons are pathologically related to Alzheimer＇s disease. Microtubule-associated protein 2 （MAP2） is of importance for neuronal, axonal, and dendritic generation, extension, and stabilization, as well as for the regulation of synaptic plasticity. OBJECTIVE： To investigate the antagonistic effects of natural-cerebrolysin-containing serum on beta amyloid protein 1-40 （Aβ1-40）-induced neurotoxicity from the standpoints of cell proliferation, synaptogenesis, and cytoskeleton formation （MAP2 expression）. DESIGN, TIME AND SETTING： A paralleled, controlled, neural cell, and molecular biology experiment was performed at the Institute of Integrated Chinese and Western Medicine, Shenzhen Hospital, Southern Medical University between February 2006 and April 2008. MATERIALS： PC12 cells, derived from the rat central nervous system, were purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China. A β1-40 was provided by Sigma, USA. Natural-cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. The natural-cerebrolysin was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yixingye （Ginkgo Leaf） in a proportion of 1：2：2. Following conventional water extraction technology, an extract （1：20） was prepared. Each gram of extract equaled 20 grams of crude drug. In a total of 12 adult male New Zealand rabbits, six underwent intragastric administration of natural-cerebrolysin extract for 1 month to prepare natural-cerebrolysin-containing serum, and the remaining six rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： An AIzheimer＇s disease in vitro model was induced in PC12 cells using Aβ1-40. The cells were incubated with varying doses of natural-cerebrolysin-containing serum （2.5%, 5%, and 10%）. Normal blank serum-treated PC12 cells served as a blank

关键词： natural-cerebrolysin Alzheimer's disease in vitro model neurotoxicity neuroprotective effect amyloid beta protein 1-40

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Toward three-dimensional in vitro models to study neurovascular unit functions in health and disease

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Neural Regeneration Research 2021年第11期16卷 2132-2140页

作者： Tara M.Caffrey Emily B.Button Jerome Robert Djavad Mowafaghian Center for Brain Health University of British ColumbiaVancouverBCCanada Department of Pathology University of British ColumbiaVancouverBCCanada Institute of Clinical Chemistry University Hospital of ZurichZurichSwitzerland

The high metabolic demands of the brain require an efficient vascular system to be coupled with neural activity to supply adequate nutrients and *** supply is coordinated by the action of neurons,glial and vascular cells,known collectively as the neurovascular unit,which temporally and spatially regulate local cerebral blood flow through a process known as neurovascular *** many neurodegenerative diseases,changes in functions of the neurovascular unit not only impair neurovascular coupling but also permeability of the blood-brain barrier,cerebral blood flow and clearance of waste from the *** order to study disease mechanisms,we need improved physiologicallyrelevant human models of the neurovascular *** towards modeling the cellular complexity of the neurovascular unit in vitro have been made using stem-cell derived organoids and more recently,vascularized organoids,enabling intricate studies of non-cell autonomous *** and design innovations in microfluidic devices and tissue engineering are progressing our ability to interrogate the *** advanced models are being used to gain a better understanding of neurodegenerative disease processes and potential *** innovation is required to build more physiologically-relevant models of the neurovascular unit encompassing both the cellular complexity and designed features to interrogate neurovascular unit functionality.

关键词： Alzheimer’s disease cerebrovasculature in vitro model neurodegeneration neurovascular unit

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IN vitro GLYCATION OF HEMOGLOBIN (HB):CHOOSING THE APPROPRIATE model

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Journal of Pharmaceutical Analysis 1995年第2期9卷 180-180页

作者：房志宇

Human HbA is nonenzymatically glycated at several sites. Approximately half of glycated Hb is formed by the addition of glucose to the amino-terminal valine of the Hb βchains, this species is called *** studies examining the effects of various agents on Hb glycation have focused on HbA1c formation in ***,approximately hal f of Hb glycation in vivo also occurs at other sites,*** amino groups on *** purpose,therefore,was to develop a model for testing the effects of several parameters and /or chemicals on glycohemoglobin formation which would be representative of the *** in vivo in terms of glycation *** was glycated by several methods:① Drying of whole blood on filter paper ② Incubation of erythrocyte hemolysates in various buffers ③Incubation of intact erythrocytes in plasma and cell culture medium. Results show that drying of Hb on filter paper caused rapid .Hb glycation which could be measured by affinity chromatography; the measured glycated Hb more than doubled in less than one week. However,measurement of HbA1c formation by immunoassay showed minimal glycation at the βchain N-terminal valine from filter paper elutes(HbA1c increased by less than 2% in one week).Similarly, incubation of hemolysates showed minimal formation of HbA1c compared to other glycation products. However,incubation of intact erythtocytes in either plasma or culture medium showed formation of HbA1c and iotal glycated Hb in proportions similar to that formed in vivo. We conclude that in vitro conditions will affect the Hb glycation site and that incubation of intact erythrocytes provides the most representative model for the study of Hb glycation.

关键词： in vitro model glycohemoglobin HbA1c

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Bioprinting of novel 3D tumor array chip for drug screening

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Bio-Design and Manufacturing 2020年第3期3卷 175-188页

作者： Mingjun Xie Qing Gao Jianzhong Fu Zichen Chen Yong He State Key Laboratory of Fluid Power and Mechatronic Systems School of Mechanical EngineeringZhejiang UniversityHangzhou 310027China Key Laboratory of 3D Printing Process and Equipment of Zhejiang Province School of Mechanical EngineeringZhejiang UniversityHangzhou 310027China

Biomedical field has been seeking a feasible standard drug screening system consisting of 3D tumor model array for drug researching due to providing sufficient samples and simulating actual in vivo tumor growth situation,which is still a challenge to rapidly and uniformly establish ***,we propose a novel drug screening system,namely 3D tumor array chip with“layer cake”structure,for drug *** gelatin methacryloyl hydrogel droplets(~0.1μL)containing tumor cells can be automatically deposited on demand with electrohydrodynamic 3D *** conductive membrane is introduced as a chip basement for preventing charges accumulation during fabricating and convenient observing during *** chambers formed by stainless steel and silicon interlayer is convenient to be assembled and *** this chip is compatible with the existing 96-well culturing plate,the drug screening protocols could keep the same as *** properties of this chip,namely printing stability,customizability,accuracy,microenvironment,tumor functionalization,are detailly *** a demonstration,it is applied for screening of epirubicin and paclitaxel with breast tumor cells to confirm the compatibility of the proposed screening system with the traditional screening *** believe this chip will potentially play a significant role in drug evaluation in the future.

关键词： 3D tumor array chip(3D-TAC) Gelatin methacryloyl(GelMA) Drug screening in vitro model Bioprinting

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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

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