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Journal of Ocean University of China 2024年第1期23卷 216-220页

作者： YU Guihong ZHOU Luning WU Guangwei LI Dehai Shandong Province Key Laboratory of Applied Mycology and Qingdao International Center on Microbes Utilizing BiogasSchool of Life SciencesQingdao Agricultural UniversityQingdao 266109China School of Medicine and Pharmacy and Sanya Oceanographic InstitutionOcean University of ChinaQingdao 266003China college of Chemical Engineering Nanjing Forestry UniversityNanjing 210037China Laboratory for Marine Drugs and Bioproducts Laoshan LaboratoryQingdao 266237China

Two new dihydropyrones,rhytismatones C(1)and D(2),and a known compound,penicillenol A1(3),were isolated from the co-culture broth of the deep-sea-derived fungus Penicillium crustosum PRB-2 and Suaeda salsa-derived endophytic fungus Peni-cillium citrinum HDN11-186.Their structures were elucidated through comprehensive analysis of nuclear magnetic resonance(NMR)spectra and mass spectra.The absolute configurations of new compounds were determined by calculating the electronic circular di-chroism(ECD)spectrum.UPLC-MS data showed that compounds 1–3 could only be detected in the media of co-culture,suggesting new biosynthetic pathways were activated in the co-cultured fungi.compound 1 showed obvious antibacterial activities against Pro-teus sp.MMBC-1002 and Bacillus subtilis MMBC-1004 with minimum inhibitory concentration(MIC)both at 25μmolL^(-1).

关键词： co-culture Penicillium crustosum Penicillium citrinum dihydropyrones antibacterial activity

Linear polyketides produced by co-culture of Penicillium crustosum and Penicillium fellutanum

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Marine Life Science & Technology 2022年第2期4卷 237-244页

作者： Guihong Yu Peng Sun Reyilamu Aierken Chunxiao Sun Zhenzhen Zhang Qian Che Guojian Zhang Tianjiao Zhu Qianqun Gu Mingyu Li Dehai Li Key Laboratory of Marine Drugs Chinese Ministry of EducationSchool of Medicine and PharmacyOcean University of ChinaQingdao266003China Laboratory for Marine Drugs and Bioproducts of Pilot National Laboratory for Marine Science and Technology(Qingdao) Qingdao266237China Fujian Provincial Key Laboratory of Innovative Drug Target Research School of Pharmaceutical ScienceXiamen UniversityXiamen361102China School of Life Sciences Shandong Province Key Laboratory of Applied MycologyQingdao Agricultural UniversityQingdao266109China

Two new polyketides, penifellutins A (1) and B (2), possessing a 22 carbon linear skeleton, were isolated from a co-culture of the deep-sea-derived fungi Penicillium crustosum PRB-2 and Penicillium fellutanum HDN14-323. Meanwhile, two esterification products of 1, penifellutins C (3) and D (4), were obtained because compound 1 could be esterified spontaneously when stored in methanol. Their configurations were difficult to determine because of chiral central crowdedness, structural flexibility and instability. As such, we solved this issue by comprehensively using Mo2(OAc)4-based CD experiments, density functional theory calculation of 13C NMR, DP4 + probability analysis and many chemical reactions, including making acetonide derivative, Mosher’s method, PGME method, etc. compounds 1 and 2 show obvious inhibitory activity on the liver hyperplasia of zebrafish larvae at a concentration of 10 μmol/L, while 3 and 4 show no activity, indicating that two carboxyls in the structure are important active sites.

关键词： co-culture Polyketides Extraction and isolation Structure elucidation Antiproliferation

Osteoblast integration of dental implant materials after challenge by sub-gingival pathogens: a co-culture study in vitro

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International Journal of Oral Science 2015年第4期7卷 250-258页

作者： Bingran Zhao Henny C van der Mei Minie Rustema-Abbing Henk J Busscher Yijin Ren Department of Orthodontics University of Groningen and University Medical Center Groningen Department of Biomedical Engineering University of Groningen and University Medical Center Groningen

Sub-gingival anaerobic pathogens can colonize an implant surface to compromise osseointegration of dental implants once the soft tissue seal around the neck of an implant is broken. In vitro evaluations of implant materials are usually done in monoculture studies involving either tissue integration or bacterial colonization. co-culture models, in which tissue cells and bacteria battle simultaneously for estate on an implant surface, have been demonstrated to provide a better in vitro mimic of the clinical situation. Here we aim to compare the surface coverage by U2OS osteoblasts cells prior to and after challenge by two anaerobic sub-gingival pathogens in a co-culture model on differently modified titanium （Ti）, titanium-zirconium （TiZr） alloys and zirconia surfaces. Monoculture studies with either U2OS osteoblasts or bacteria were also carried out and indicated significant differences in biofilm formation between the implant materials, but interactions with U2OS osteoblasts were favourable on all materials. Adhering U2OS osteoblasts cells, however, were significantly more displaced from differently modified Ti surfaces by challenging sub-gingival pathogens than from TiZr alloys and zirconia variants. combined with previous work employing a co-culture model consisting of human gingival fibroblasts and supra-gingival oral bacteria, results point to a different material selection to stimulate the formation of a soft tissue seal as compared to preservation of osseointegration under the unsterile conditions of the oral cavity.

关键词： biofilm co-culture dental implant osteoblasts sub-gingival pathogens titanium-zirconium alloy

In vitro chondrogenesis of the goat bone marrow mesenchymal stem cells directed by chondrocytes in monolayer and 3-dimetional indirect co-culture system

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Chinese Medical Journal 2011年第19期124卷 3080-3086页

作者： LI Jian-wei GUO Xiao-lei HE Chun-la TUO Yong-hua WANG Zhao WEN Jun JIN Dan Department of Traumatology and Orthopedics Nanfang Hospital Southern Medical University Guangzhou Guangdong 510515 China Department of Biostatistics Southern Medical University Guangzhu Gungdong 510515 China

Background Cartilage injury has a very poor capacity for intrinsic regeneration. The cell-based treatment strategy for the cartilage repair using differentiated bone marrow mesenchymal stem cells （BMSCs） is, however, a promising approach to the chondral repair. This study was aimed to explore the chondrogenic potential of the goat BMSCs in the Transwell co-culture system and the poly-laetide-co-glycolide （PLGA） scaffolds.Methods The BMSCs were isolated from the goat iliac crest while the chondrocytes were obtained from the goat＇s last costal cartilage. In the Transwell co-culture system, the BMSCs co-cultured with chondrocytes were designed as group A,whereas the goat＇s BMSCs induced with the chondrogenic medium were group B. Both groups A and B were the experimental groups, while group C that only contained BMSCs was the control group. In the PLGA scaffolds co-culture system, BMSCs were seeded into the PLGA scaffolds, which were suspended in the 24-well plate, and the control group was established by presence or absence of chondrocytes at the bottom of the 24-well plate. Toluidine blue staining,Alcian blue staining, collagen Ⅱ immunofluoresence, collagen Ⅱ immunochemical staining, collagen Ⅰ, collagen Ⅱ, coL2a Q-PCR and osteopontin Q-PCR were used to examine the chondrogenic conditions as well as the expressions of chondrogenic and osteogenic genes.Results Cells isolated from the aspirates of the goat bone marrow proliferated rapidly and gained characteristics of stem cells in Passage 4. However, the differentiations of chondrocytes were not apparent in Passage 3. The results from Toluidine blue staining, collagen Ⅱ immunofluoresence and PCR showed the transformation of BMSCs to chondrocytes in the Transwell co-culture system and PLGA scaffolds. Although the cartilage gene expressions were upgraded in both chondrogenesis group and co-culture system, the osteopontin gene expression, which represents osteogenic level, was also up-regulated.conclusions The Transwell co-culture system and the PLGA scaffolds co-cultu

关键词： bone marrow mesenchymal stem cells chondrocytes co-culture goat

Investigating materials and orientation parameters for the creation of a 3D musculoskeletal interface co-culture model

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Regenerative Biomaterials 2020年第4期7卷 413-425页

作者： Hamad Alsaykhan Jennifer Z.Paxton Anatomy@Edinburgh Edinburgh Medical School:Biomedical SciencesThe University of EdinburghTeviot PlaceEdinburgh EH89AGUK Department of Basic Medical Sciences College of Medicine and Health SciencesQassim UniversityPO Box 99151911 Unaizah CampusAl-Qassim 51911Saudi Arabia

Musculoskeletal tissue interfaces are a common site of injury in the young,active populations.In particular,the interface between the musculoskeletal tissues of tendon and bone is often injured and to date,no single treatment has been able to restore the form and function of damaged tissue at the bone-tendon interface.Tissue engineering and regeneration hold great promise for the manufacture of bespoke in vitro models or implants to be used to advance repair and so this study investigated the material,orientation and culture choices for manufacturing a reproducible 3D model of a musculoskeletal interface between tendon and bone cell populations.Such models are essential for future studies focussing on the regeneration of musculoskeletal interfaces in vitro.Cell-encapsulated fibrin hydrogels,arranged in a horizontal orientation though a simple moulding procedure,were shown to best support cellular growth and migration of cells to form an in vitro tendon-bone interface.This study highlights the importance of acknowledging the material and technical challenges in establishing co-cultures and suggests a reproducible methodology to form 3D co-cultures between tendon and bone,or other musculoskeletal cell types,in vitro.

关键词： tendon bone 3D co-culture hydrogels musculoskeletal interface

In vitro model of the blood-brain barrier established by co-culture of primary cerebral microvascular endothelial and astrocyte cells

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Neural Regeneration Research 2015年第12期10卷 2011-2017页

作者： Yan Wang Ning Wang Biao Cai Guang-yun Wang Jing Li Xing-xing Piao Key Laboratory of Xin’an Medicine Ministry of Education Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine Anhui Academy of Chinese college of Pharmacy Anhui University of Chinese Medicine

Drugs for the treatment and prevention of nervous system diseases must permeate the bloodbrain barrier to take effect.In vitro models of the blood-brain barrier are therefore important in the investigation of drug permeation mechanisms.However,to date,no unified method has been described for establishing a blood-brain barrier model.Here,we modified an in vitro model of the blood-brain barrier by seeding brain microvascular endothelial cells and astrocytes from newborn rats on a polyester Transwell cell culture membrane with 0.4-μm pores,and conducted transepithelial electrical resistance measurements,leakage tests and assays for specific bloodbrain barrier enzymes.We show that the permeability of our model is as low as that of the bloodbrain barrier in vivo.Our model will be a valuable tool in the study of the mechanisms of action of neuroprotective drugs.

关键词： nerve regeneration blood-brain barrier astrocytes brain microvascular endothelial cells permeability co-culture Transwell chamber neural regeneration

co-culture:stimulate the metabolic potential and explore the molecular diversity of natural products from microorganisms

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Marine Life Science & Technology 2021年第3期3卷 363-374页

作者： Xiao-Yue Peng Jin-Tao Wu Chang-Lun Shao Zhi-Yong Li Min Chen Chang-Yun Wan Key Laboratory of Marine Drugs The Ministry of Education of ChinaSchool of Medicine and PharmacyInstitute of Evolution and Marine BiodiversityOcean University of ChinaQingdao266003China Marine Science and Technology Institute College of Environmental Science and EngineeringYangzhou UniversityYangzhou225127China Laboratory for Marine Drugs and Bioproducts Qingdao National Laboratory for Marine Science and TechnologyQingdao266237China State Key Laboratory of Microbial Metabolism School of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai201100China

Microbial secondary metabolites have long been considered as potential sources of lead compounds for medicinal use due to their rich chemical diversity and extensive biological activities.However,many biosynthetic gene clusters remain silent under traditional laboratory culture conditions,resulting in repeated isolation of a large number of known compounds.The co-culture strategy simulates the complex ecological environment of microbial life by using an ecology-driven method to activate silent gene clusters of microorganisms and tap their metabolic potential to obtain novel bioactive secondary metabolites.In this review,representative studies from 2017 to 2020 on the discovery of novel bioactive natural products from co-cultured microorganisms are summarized.A series of natural products with diverse and novel structures have been discovered successfully by co-culture strategies,including fungus-fungus,fungus-bacterium,and bacterium-bacterium co-culture approaches.These novel compounds exhibited various bioactivities including extensive antimicrobial activities and potential cytotoxic activities,especially when it came to disparate marine-derived species and cross-species of marine strains and terrestrial strains.It could be concluded that co-culture can be an effective strategy to tap the metabolic potential of microorganisms,particularly for marine-derived species,thus providing diverse molecules for the discovery of lead compounds and drug candidates.

关键词： co-culture Microorganisms Secondary metabolites Chemical diversity

co-culture of Schwann cells and endothelial cells for synergistically regulating dorsal root ganglion behavior on chitosan-based anisotropic topology for peripheral nerve regeneration

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Burns & Trauma 2022年第1期10卷 340-356页

作者： Tiantian Zheng Linliang Wu Shaolan Sun Jiawei Xu Qi Han Yifan Liu Ronghua Wu Guicai Li Key laboratory of Neuroregeneration of Jiangsu and Ministry of Education Co-innovation Center of Neuroregenera-tionNMPA Key Lab for Research and Evaluation of Tissue Engineering Technology ProductsNantong University.226001NantongP.R.China School of Medicine Nantong University.226001NantongP.R.China Guangxi Key Laboratory of Regenerative Medicine Guangxi Medical University530021NanningP.R.China National Engineering Laboratory for Modern Silk Soochow UniversitySuzhou 215123China

Background:Anisotropic topologies are known to regulate cell-oriented growth and induce cell differentiation,which is conducive to accelerating nerve regeneration,while co-culture of endothelial cells(ECs)and Schwann cells(SCs)can significantly promote the axon growth of dorsal root ganglion(DRG).However,the synergistic regulation of EC and SC co-culture of DRG behavior on anisotropic topologies is still rarely reported.The study aims to investigate the effect of anisotropic topology co-cultured with Schwann cells and endothelial cells on dorsal root ganglion behavior for promoting peripheral nerve regeneration.Methods:Chitosan/artemisia sphaerocephala(CS/AS)scaffolds with anisotropic topology were first prepared using micro-molding technology,and then the surface was modified with dopamine to facilitate cell adhesion and growth.The physical and chemical properties of the scaffolds were characterized through morphology,wettability,surface roughness and component variation.SCs and ECs were co-cultured with DRG cells on anisotropic topology scaffolds to evaluate the axon growth behavior.Results:Dopamine-modified topological CS/AS scaffolds had good hydrophilicity and provided an appropriate environment for cell growth.Cellular immunofluorescence showed that in contrast to DRG growth alone,co-culture of SCs and ECs could not only promote the growth of DRG axons,but also offered a stronger guidance for orientation growth of neurons,which could effectively prevent axons from tangling and knotting,and thus may significantly inhibit neurofibroma formation.Moreover,the co-culture of SCs and ECs could promote the release of nerve growth factor and vascular endothelial growth factor,and up-regulate genes relevant to cell proliferation,myelination and skeletal development via the PI3K-Akt,MAPK and cytokine and receptor chemokine pathways.conclusions:The co-culture of SCs and ECs significantly improved the growth behavior of DRG on anisotropic topological scaffolds,which may provide an important basis for the development of

关键词： Schwann cells Endothelial cells Dorsal root ganglion behavior co-culture Anisotropic topology Regulation mechanism Nerve Regeneration

Osteogenic Differentiation of Bone Mesenchymal Stem Cells Regulated by Osteoblasts under EMF Exposure in a co-culture System

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2014年第2期34卷 247-253页

作者：虞冀哲吴华杨勇刘朝旭刘阳宋明宇 Department of Orthopedics Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

This study examined the osteogenic effect of electromagnetic fields （EMF） under the simulated in vivo conditions. Rat bone marrow mesenchymal stem cells （BMSCs） and rat osteoblasts were co-cultured and exposed to 50 Hz, 1.0 mT EMF for different terms. Unexposed single-cultured BMSCs and osteoblasts were set as controls. Cell proliferation features of single-cultured BMSCs and osteoblasts were studied by using a cell counting kit （CCK-8）. For the co-culture system, cells in each group were randomly chosen for alkaline phosphatase （ALP） staining on the day 7. When EMF exposure lasted for 14 days, dishes in each group were randomly chosen for total RNA extraction and von Kossa staining. The mRNA expression of osteogenic markers was detected by using real-time PCR. Our study showed that short-term EMF exposure （2 h/day） could obviously promote prolifera- tion of BMSCs and osteoblasts, while long-term EMF （8 h/day） could promote osteogenic differen- tiation significantly under co-cultured conditions. Under EMF exposure, osteogenesis-related mRNA expression changed obviously in co-cultured and single-cultured cells. It was noteworthy that most osteogenic indices in osteoblasts were increased markedly after co-culture except Bmp2, which was increased gradually when ceils were exposed to EMF. compared to other indices, the expression of Bmp2 in BMSCs was increased sharply in both single-cultured and co-cultured groups when they were exposed to EMF. The mRNA expression of Bmp2 in BMSCs was approximately four times higher in 8-h EMF group than that in the unexposed group. Our results suggest that Bmp2-mediated cellular interaction induced by EMF exposure might play an important role in the osteogenic differ- entiation of BMSCs.

关键词： electromagnetic fields bone marrow mesenchymal stem cell osteoblast osteogenicmechanism co-culture

co-culture of Mesenchymal Stem Cells with Umbilical Vein Endothelial Cells under Hypoxic condition

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2012年第2期32卷 173-180页

作者：张波杨述华张宇坤孙志博许伟华叶树楠 Department of Orthopedics Union Hospital Tongji Medical College Huazhong University of Science and Technology

By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein endothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of transplanted hMSCs for the treatment of avascular ni ANFH, the effect of hMSCs on survival, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition were investigated in vitro. hMSCs and HUVECs were cultured and identified in vitro. Three kinds of conditioned media, CdM-CdMNOR, CdM-CdMHYP and HUVEC-CdMHYP were prepared. HUVECs were cultured with these conditioned media under hypoxia. The survival rate, apoptosis rate, migration and angiogenesis of HUVECs were respectively detected by CCK-8, flow cytometry, Transwell and tube formation assay. The content of SDF-1α, VEGF and IL-6 in CdM was determined by ELISA. Our results showed that hMSCs and HUVECs were cultured and identified successfully. compared with MSC-CdMNOR and HUVEC-CdMHYP groups, the survival rate, migra-tion and angiogenesis of HUVECs in MSC-CdMHYP group were significantly increased while the apoptosis rate was declined (Pco-culture of hMSCs and HUVECs. The underlying mechanism may be that hMSCs could secrete a number of cytokines and improve niche, which might be helpful in the treatment of femoral head necrosis.

关键词： hypoxia mesenchymal stem cells umbilical vein endothelial cells co-culture femoral head necrosis