Ex vivo lung perfusion(EVLP) is a powerful experimental model for isolated lung research. EVLP allows for the lungs to be manipulated and characterized in an external environment so that the effect of specific ventila...Ex vivo lung perfusion(EVLP) is a powerful experimental model for isolated lung research. EVLP allows for the lungs to be manipulated and characterized in an external environment so that the effect of specific ventilation/perfusion variables can be studied independent of other confounding physiologic contributions. At the same time,EVLP allows for normal organ level function and real-time monitoring of pulmonary physiology and mechanics. As a result,this technique provides uniqueadvantages over in vivo and in vitro models. Small and large animal models of EVLP have been developed and each of these models has their strengths and weaknesses. In this manuscript,we provide insight into the relative strengths of each model and describe how the development of advanced EVLP protocols is leading to a novel experimental platform that can be used to answer critical questions in pulmonary physiology and transplant medicine.展开更多
Objective:Using neuromyelitis optica immunoglobulin G(NMO-IgG)to induced ex vivo mice spinal cord slice model.Methods:Vibratome-cut transverse spinal cord slices from 7-day-old C57BL/6Jmouse pups were cultured on tran...Objective:Using neuromyelitis optica immunoglobulin G(NMO-IgG)to induced ex vivo mice spinal cord slice model.Methods:Vibratome-cut transverse spinal cord slices from 7-day-old C57BL/6Jmouse pups were cultured on transwell porous supports for 7days,then randomly divided into the control group and NMO model group.Slices of the control group were further cultured with human serum complement,while slices from NMO model group were exposed to complement and NMO-IgG.After 24-hour incubation,slices of both groups were measured for aquaporin-4(AQP4),glial fibrillary acidic protein(GFAP),myelin basic protein(MBP)and neurofilament light chain(NFL)by immunofluorescence.Results:Slices exposed to NMO-IgG showed astrocyte swelling,and a significant loss of AQP4and GFAP staining.Ratios of the loss of AQP4and GFAP staining were 77.74%±6.75%and 75.62%±5.76%respectively in the model group,and NMO-like injury score was 3.11±0.60.But there were no obvious losses of AQP4and GFAP staining in the control group,and NMO-like lesion score was 0.00.There were significant differences between the two groups with regards to the above indexes(P<0.01).Ratios of the loss of MBP and NFL staining in the model group were 37.60%±4.88%and46.29%±4.98%respectively,while the corresponding figures in the control group were 9.10%±1.63%and 5.80%±0.81%,and the differences between the two groups were statistically significant(P<0.01).Conclusion:These results suggested that NMO-IgG-induced ex vivo spinal cord slice model possesses typical features of NMO,and this model might be useful for relevant fundamental studies.展开更多
Efforts to find an ideal model for pancreatitis date back to 1960’s. Many models are suggested since then. Every model has its own advantages and disadvantages. Some of these models test etiology while others simulat...Efforts to find an ideal model for pancreatitis date back to 1960’s. Many models are suggested since then. Every model has its own advantages and disadvantages. Some of these models test etiology while others simulate the complications of pancreatitis. An ideal model which by itself demonstrates all aspects of pancreatitis including systemic changes is yet to be described. In this review we tried to gather the basic, easy to construct models.展开更多
BACKGROUND Liver fibrosis can result in end-stage liver failure and death.AIM To examine human liver fibrogenesis and anti-fibrotic therapies,we evaluated the three dimensional ex vivo liver slice(LS)model.METHODS Fib...BACKGROUND Liver fibrosis can result in end-stage liver failure and death.AIM To examine human liver fibrogenesis and anti-fibrotic therapies,we evaluated the three dimensional ex vivo liver slice(LS)model.METHODS Fibrotic liver samples(F0 to F4 fibrosis stage according to the METAVIR score)were collected from patients after liver resection.Human liver slices(HLS)were cultivated for up to 21 days.Hepatitis C virus(HCV)infection,alcohol(ethanol stimulation)and steatosis(palmitate stimulation)were examined in fibrotic(F2 to F4)liver slices infected(or not)with HCV.F0-F1 HLS were used as controls.At day 0,either ursodeoxycholic acid(choleretic and hepatoprotective properties)and/or α-tocopherol(antioxidant properties)were added to standard of care on HLS and fibrotic liver slices,infected(or not)with HCV.Expression of the biomarkers of fibrosis and the triglyceride production were checked by quantitative reverse transcription polymerase chain reaction and/or enzymelinked immunosorbent assay.RESULTS The cultures were viable in vitro for 21 days allowing to study fibrosis inducers and to estimate the effect of anti-fibrotic drugs.Expression of the biomarkers of fibrosis and the progression to steatosis(estimated by triglycerides production)was increased with the addition of HCV and/or ethanol or palmitate.From day 15 of the follow-up studies,a significant decrease of both transforming growth factorβ-1 and Procol1A1 expression and triglycerides production was observed when a combined anti-fibrotic treatment was applied on HCV infected F2-F4 LS cultures.CONCLUSION These results show that the human three dimensional ex vivo model effectively reflects the in vivo processes in damaged human liver(viral,alcoholic,nonalcoholic steatohepatitis liver diseases)and provides the proof of concept that the LS examined model permits a rapid evaluation of new anti-fibrotic therapies when used alone or in combination.展开更多
Skin is widely used as a drug delivery route due to its easy access and the possibility of using relatively painless methods for the administration of bioactive molecules.However,the barrier properties of the skin,alo...Skin is widely used as a drug delivery route due to its easy access and the possibility of using relatively painless methods for the administration of bioactive molecules.However,the barrier properties of the skin,along with its multilayer structure,impose severe restrictions on drug transport and bioavailability.Thus,bioengineered models aimed at emulating the skin have been developed not only for optimizing the transdermal transport of different drugs and testing the safety and toxicity of substances but also for understanding the biological processes behind skin wounds.Even though in vivo research is often preferred to study biological processes involving the skin,in vitro and ex vivo strategies have been gaining increasing relevance in recent years.Indeed,there is a noticeably increasing adoption of in vitro and ex vivo methods by internationally accepted guidelines.Furthermore,microfluidic organ-on-a-chip devices are nowadays emerging as valuable tools for functional and behavioural skin emulation.Challenges in miniaturization,automation and reliability still need to be addressed in order to create skin models that can predict skin behaviour in a robust,high-throughput manner,while being compliant with regulatory issues,standards and guidelines.In this review,skin models for transdermal transport,wound repair and cutaneous toxicity will be discussed with a focus on high-throughput strategies.Novel microfluidic strategies driven by advancements in microfabrication technologies will also be revised as a way to improve the efficiency of existing models,both in terms of complexity and throughput.展开更多
Hepatitis C virus(HCV) is a major cause of liver diseases including liver cirrhosis and hepatocellular carcinoma. Approximately 3% of the world population is infected with HCV. Thus, HCV infection is considered a publ...Hepatitis C virus(HCV) is a major cause of liver diseases including liver cirrhosis and hepatocellular carcinoma. Approximately 3% of the world population is infected with HCV. Thus, HCV infection is considered a public healthy challenge. It is worth mentioning, that the HCV prevalence is dependent on the countries with infection rates around 20% in high endemic countries. The review summarizes recent data on HCV molecular biology, the physiopathology of infection(immune-mediated liver damage, liver fibrosis and lipid metabolism), virus diagnostic and treatment. In addition, currently available in vitro, ex vivo and animal models to study the virus life cycle, virus pathogenesis and therapy are described. Understanding of both host and viral factors may in the future lead to creation of new approaches in generation of an efficient therapeutic vaccine.展开更多
文摘Ex vivo lung perfusion(EVLP) is a powerful experimental model for isolated lung research. EVLP allows for the lungs to be manipulated and characterized in an external environment so that the effect of specific ventilation/perfusion variables can be studied independent of other confounding physiologic contributions. At the same time,EVLP allows for normal organ level function and real-time monitoring of pulmonary physiology and mechanics. As a result,this technique provides uniqueadvantages over in vivo and in vitro models. Small and large animal models of EVLP have been developed and each of these models has their strengths and weaknesses. In this manuscript,we provide insight into the relative strengths of each model and describe how the development of advanced EVLP protocols is leading to a novel experimental platform that can be used to answer critical questions in pulmonary physiology and transplant medicine.
基金supported by the National Natural Science Foundation of China (No.81460194 No.81260188)
文摘Objective:Using neuromyelitis optica immunoglobulin G(NMO-IgG)to induced ex vivo mice spinal cord slice model.Methods:Vibratome-cut transverse spinal cord slices from 7-day-old C57BL/6Jmouse pups were cultured on transwell porous supports for 7days,then randomly divided into the control group and NMO model group.Slices of the control group were further cultured with human serum complement,while slices from NMO model group were exposed to complement and NMO-IgG.After 24-hour incubation,slices of both groups were measured for aquaporin-4(AQP4),glial fibrillary acidic protein(GFAP),myelin basic protein(MBP)and neurofilament light chain(NFL)by immunofluorescence.Results:Slices exposed to NMO-IgG showed astrocyte swelling,and a significant loss of AQP4and GFAP staining.Ratios of the loss of AQP4and GFAP staining were 77.74%±6.75%and 75.62%±5.76%respectively in the model group,and NMO-like injury score was 3.11±0.60.But there were no obvious losses of AQP4and GFAP staining in the control group,and NMO-like lesion score was 0.00.There were significant differences between the two groups with regards to the above indexes(P<0.01).Ratios of the loss of MBP and NFL staining in the model group were 37.60%±4.88%and46.29%±4.98%respectively,while the corresponding figures in the control group were 9.10%±1.63%and 5.80%±0.81%,and the differences between the two groups were statistically significant(P<0.01).Conclusion:These results suggested that NMO-IgG-induced ex vivo spinal cord slice model possesses typical features of NMO,and this model might be useful for relevant fundamental studies.
文摘Efforts to find an ideal model for pancreatitis date back to 1960’s. Many models are suggested since then. Every model has its own advantages and disadvantages. Some of these models test etiology while others simulate the complications of pancreatitis. An ideal model which by itself demonstrates all aspects of pancreatitis including systemic changes is yet to be described. In this review we tried to gather the basic, easy to construct models.
基金the Institut National de la Santéet de la Recherche Médicale(INSERM,France)and by Institut Pasteur(Paris,France)Daria Kartasheva-Ebertz received a PhD Fellowship from Assistance Publique-Hôpitaux de Paris(APHP,France).
文摘BACKGROUND Liver fibrosis can result in end-stage liver failure and death.AIM To examine human liver fibrogenesis and anti-fibrotic therapies,we evaluated the three dimensional ex vivo liver slice(LS)model.METHODS Fibrotic liver samples(F0 to F4 fibrosis stage according to the METAVIR score)were collected from patients after liver resection.Human liver slices(HLS)were cultivated for up to 21 days.Hepatitis C virus(HCV)infection,alcohol(ethanol stimulation)and steatosis(palmitate stimulation)were examined in fibrotic(F2 to F4)liver slices infected(or not)with HCV.F0-F1 HLS were used as controls.At day 0,either ursodeoxycholic acid(choleretic and hepatoprotective properties)and/or α-tocopherol(antioxidant properties)were added to standard of care on HLS and fibrotic liver slices,infected(or not)with HCV.Expression of the biomarkers of fibrosis and the triglyceride production were checked by quantitative reverse transcription polymerase chain reaction and/or enzymelinked immunosorbent assay.RESULTS The cultures were viable in vitro for 21 days allowing to study fibrosis inducers and to estimate the effect of anti-fibrotic drugs.Expression of the biomarkers of fibrosis and the progression to steatosis(estimated by triglycerides production)was increased with the addition of HCV and/or ethanol or palmitate.From day 15 of the follow-up studies,a significant decrease of both transforming growth factorβ-1 and Procol1A1 expression and triglycerides production was observed when a combined anti-fibrotic treatment was applied on HCV infected F2-F4 LS cultures.CONCLUSION These results show that the human three dimensional ex vivo model effectively reflects the in vivo processes in damaged human liver(viral,alcoholic,nonalcoholic steatohepatitis liver diseases)and provides the proof of concept that the LS examined model permits a rapid evaluation of new anti-fibrotic therapies when used alone or in combination.
基金supported by the grants from Natural Science Foundation of Beijing(No.7042055)from the Basic Research Program of the Ministry of Science and Technology(No.2013FY113800)~~
基金supported by Portuguese funds from the Foundation for Science and Technology(FCT)(projects i3S,ref.UID/BIM/04293/2020,SKINCHIP,ref.PTDC/BBBBIO/1889/2014,iMed.ULisboa,refs.UID/DTP/04138/2020 and UIDB/DTP/04138/2020)UK funds from the Engineering and Physical Sciences Research Council(EPSRC)and Medical Research Council(MRC)(projects Centre for Doctoral Training in Regenerative Medicine,ref.EP/L014904/1,and Henry Royce Institute for Advanced Materials,refs.EP/R00661X/1,EP/P025021/1 and EP/P025498/1).
文摘Skin is widely used as a drug delivery route due to its easy access and the possibility of using relatively painless methods for the administration of bioactive molecules.However,the barrier properties of the skin,along with its multilayer structure,impose severe restrictions on drug transport and bioavailability.Thus,bioengineered models aimed at emulating the skin have been developed not only for optimizing the transdermal transport of different drugs and testing the safety and toxicity of substances but also for understanding the biological processes behind skin wounds.Even though in vivo research is often preferred to study biological processes involving the skin,in vitro and ex vivo strategies have been gaining increasing relevance in recent years.Indeed,there is a noticeably increasing adoption of in vitro and ex vivo methods by internationally accepted guidelines.Furthermore,microfluidic organ-on-a-chip devices are nowadays emerging as valuable tools for functional and behavioural skin emulation.Challenges in miniaturization,automation and reliability still need to be addressed in order to create skin models that can predict skin behaviour in a robust,high-throughput manner,while being compliant with regulatory issues,standards and guidelines.In this review,skin models for transdermal transport,wound repair and cutaneous toxicity will be discussed with a focus on high-throughput strategies.Novel microfluidic strategies driven by advancements in microfabrication technologies will also be revised as a way to improve the efficiency of existing models,both in terms of complexity and throughput.
文摘Hepatitis C virus(HCV) is a major cause of liver diseases including liver cirrhosis and hepatocellular carcinoma. Approximately 3% of the world population is infected with HCV. Thus, HCV infection is considered a public healthy challenge. It is worth mentioning, that the HCV prevalence is dependent on the countries with infection rates around 20% in high endemic countries. The review summarizes recent data on HCV molecular biology, the physiopathology of infection(immune-mediated liver damage, liver fibrosis and lipid metabolism), virus diagnostic and treatment. In addition, currently available in vitro, ex vivo and animal models to study the virus life cycle, virus pathogenesis and therapy are described. Understanding of both host and viral factors may in the future lead to creation of new approaches in generation of an efficient therapeutic vaccine.
