AIM To develop a culture mode providingdurable biomaterials with high yields andactivities used in bioartificial liver.METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion...AIM To develop a culture mode providingdurable biomaterials with high yields andactivities used in bioartificial liver.METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion with collagenase.In culture onmicrocarriers,primary porcine hepatocyteswere inoculated at a concentration of 5×10~7/mLinto the static culture systems containing 2 g/LCytodex-3,then supplemented with 100 mL/Lfetal calf serum(FCS)or 100 mL/L porcineportal vein serum(PPVS)respectively.Inspheroidal aggregate culture hepatocytes wereinoculated into 100 mL siliconized flasks at aconcentration of 5.0×10~6/mL.RESULTS In culture on microcarriershepatocytes tended to aggregate on Cytodex-3obviously after being inoculated.Typical multi-cellular aggregated spheroids could be found inthe two systems 24 h-48 h after hepatocyteswere cultured.The morphological charact-eristics and synthetic functions were maintainedfor 5 wk in FCS culture system and 8 wk in PPVSculture system.In spheroidal aggregate cultureabout 80%-90% isolated hepatocytes becameaggregated spheroids 24h after cultured insuspension and mean diameter of the spheroidswas 100μm.The relationship among thehepatocytes resembled that in the liver in vivo.Synthetic functions of albumin and urea of the spheroids were twice those of hepatocytescultured on monolayers.CONCLUSION As high-yields and high-activitymodes of culture on microcarriers or inspheroidal aggregate culture with portal veinserum are promising to provide biomaterials forbioartificial liver(BAL)efficiently.展开更多
A large amount of propellant materials are produced every year,and storage and disposal of these propellant materials seriously contributes to environmental pollution.Alkyne terminated polybutadiene with urethane segm...A large amount of propellant materials are produced every year,and storage and disposal of these propellant materials seriously contributes to environmental pollution.Alkyne terminated polybutadiene with urethane segments(PUPB)is the macromolecule backbone of these propellant materials,and degradation of PUPB is central to the eco-friendly treatment of propellant materials.In this study,we isolated a polyurethane(PU)-and PUPB-degrading fungus from soils contaminated with rocket propellant,and the fungus H14 was identified as Fusarium solani(Mart.)Sacc.based on macro-and micro-morphology as well as phylogenetic analyses.The ability of F.solani H14 to degrade PU film and PUPB patches was evaluated via mass loss,scanning electron microscopy(SEM)and enzyme production ability.Mass loss analyses revealed a 25.8%reduction in mass of PU and 1.3%reduction in mass of PUPB after F.solani H14 was incubated with PU and PUPB for 90 days,respectively.We found that F.solani H14 mycelia significantly colonized both PU and PUPB.SEM images showed that the surface of PU films and PUPB patches formed holes,underwent folding and experienced damage as well as irregular fissuring from the erosion of fungal hypha.Moreover,two possible degradative enzymes,lipase and esterase,were produced by F.solani.Our study opens a new avenue of research for eco-friendly treatments of explosive materials and propellants.This paper represents the first article on the degradation of PUPB patches.展开更多
AIM To improve the Cultivation efficiency and yield of human liver cell line CL--l.METHODS High--density cultivation of CL--l onmicrocarriers was carried out with periodicobservation of their growth and proliferation....AIM To improve the Cultivation efficiency and yield of human liver cell line CL--l.METHODS High--density cultivation of CL--l onmicrocarriers was carried out with periodicobservation of their growth and proliferation.The specific functions of human liver cell werealso determined.RESULTS Cells of CL-1 cell line grew well onmicrocarrier Cytodex--3 and on the 7th day thepeak was reached. The amount of CLI cells was2.13 x 10’ and the total amount of albuminsynthesis reached 71 .23 ig, urea synthesis23 .32 mg and diazepam transformation 619 .7 ugrespectively. The yield of CL-l on microcarrierswas 49.3 times that of conventional cultivation.The amounts of albumin synthesis, ureasynthesis and diazepam transformation were39.8 times, 41 .6 times and 33.3 times those ofconventional Cultivation, respectively.CONCLUSION The human liver cell line CL--1 canbe Cultivated to a high density with Cytodex--3and has better biological functions. High-densityCultivation of CLI on microcarriers can act asthe biological material of bioartificial liver.展开更多
基金the National Natural Science Foundation of China,No.39570212
文摘AIM To develop a culture mode providingdurable biomaterials with high yields andactivities used in bioartificial liver.METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion with collagenase.In culture onmicrocarriers,primary porcine hepatocyteswere inoculated at a concentration of 5×10~7/mLinto the static culture systems containing 2 g/LCytodex-3,then supplemented with 100 mL/Lfetal calf serum(FCS)or 100 mL/L porcineportal vein serum(PPVS)respectively.Inspheroidal aggregate culture hepatocytes wereinoculated into 100 mL siliconized flasks at aconcentration of 5.0×10~6/mL.RESULTS In culture on microcarriershepatocytes tended to aggregate on Cytodex-3obviously after being inoculated.Typical multi-cellular aggregated spheroids could be found inthe two systems 24 h-48 h after hepatocyteswere cultured.The morphological charact-eristics and synthetic functions were maintainedfor 5 wk in FCS culture system and 8 wk in PPVSculture system.In spheroidal aggregate cultureabout 80%-90% isolated hepatocytes becameaggregated spheroids 24h after cultured insuspension and mean diameter of the spheroidswas 100μm.The relationship among thehepatocytes resembled that in the liver in vivo.Synthetic functions of albumin and urea of the spheroids were twice those of hepatocytescultured on monolayers.CONCLUSION As high-yields and high-activitymodes of culture on microcarriers or inspheroidal aggregate culture with portal veinserum are promising to provide biomaterials forbioartificial liver(BAL)efficiently.
基金This work was financed by Open Research Fund Program of Science and Technology on Aerospace Chemical Power Laboratory(STACPL320181B04)We also would like to thank the support from the National Natural Science Foundation of China(NSFC21975066,NSFC21875061).
文摘A large amount of propellant materials are produced every year,and storage and disposal of these propellant materials seriously contributes to environmental pollution.Alkyne terminated polybutadiene with urethane segments(PUPB)is the macromolecule backbone of these propellant materials,and degradation of PUPB is central to the eco-friendly treatment of propellant materials.In this study,we isolated a polyurethane(PU)-and PUPB-degrading fungus from soils contaminated with rocket propellant,and the fungus H14 was identified as Fusarium solani(Mart.)Sacc.based on macro-and micro-morphology as well as phylogenetic analyses.The ability of F.solani H14 to degrade PU film and PUPB patches was evaluated via mass loss,scanning electron microscopy(SEM)and enzyme production ability.Mass loss analyses revealed a 25.8%reduction in mass of PU and 1.3%reduction in mass of PUPB after F.solani H14 was incubated with PU and PUPB for 90 days,respectively.We found that F.solani H14 mycelia significantly colonized both PU and PUPB.SEM images showed that the surface of PU films and PUPB patches formed holes,underwent folding and experienced damage as well as irregular fissuring from the erosion of fungal hypha.Moreover,two possible degradative enzymes,lipase and esterase,were produced by F.solani.Our study opens a new avenue of research for eco-friendly treatments of explosive materials and propellants.This paper represents the first article on the degradation of PUPB patches.
基金Supported by the National Natural Science Foundation of China,No.39570212.
文摘AIM To improve the Cultivation efficiency and yield of human liver cell line CL--l.METHODS High--density cultivation of CL--l onmicrocarriers was carried out with periodicobservation of their growth and proliferation.The specific functions of human liver cell werealso determined.RESULTS Cells of CL-1 cell line grew well onmicrocarrier Cytodex--3 and on the 7th day thepeak was reached. The amount of CLI cells was2.13 x 10’ and the total amount of albuminsynthesis reached 71 .23 ig, urea synthesis23 .32 mg and diazepam transformation 619 .7 ugrespectively. The yield of CL-l on microcarrierswas 49.3 times that of conventional cultivation.The amounts of albumin synthesis, ureasynthesis and diazepam transformation were39.8 times, 41 .6 times and 33.3 times those ofconventional Cultivation, respectively.CONCLUSION The human liver cell line CL--1 canbe Cultivated to a high density with Cytodex--3and has better biological functions. High-densityCultivation of CLI on microcarriers can act asthe biological material of bioartificial liver.
