One of the greatest impacts on in vitro cell biology was the introduction of three-dimensional(3D)culture systems more than six decades ago and this era may be called the dawn of 3D-tissue culture.Although the advanta...One of the greatest impacts on in vitro cell biology was the introduction of three-dimensional(3D)culture systems more than six decades ago and this era may be called the dawn of 3D-tissue culture.Although the advantages were obvious,this field of research was a "sleeping beauty"until the 1970s when multicellular spheroids were discovered as ideal tumor models.With this rebirth,organotypical culture systems became valu-able tools and this trend continues to increase.While in the beginning,simple approaches,such as aggregation culture techniques,were favored due to their simplicity and convenience,now more sophisticated systems are used and are still being developed.One of the boosts in the development of new culture techniques arises from elaborate manufacturing and surface modification tech-niques,especially micro and nano system technologies that have either improved dramatically or have evolved very recently.With the help of these tools,it will soon be possible to generate even more sophisticated and more organotypic-like culture systems.Since 3D per-fused or superfused systems are much more complex to set up and maintain compared to use of petri dishes and culture flasks,the added value of 3D approaches still needs to be demonstrated.展开更多
Previous studies have proved that dynamic culture could facilitate nutrients transport and apply mechanical stimulation to the cells within three-dimensional scaffolds,thus enhancing the differentiation of stem cells ...Previous studies have proved that dynamic culture could facilitate nutrients transport and apply mechanical stimulation to the cells within three-dimensional scaffolds,thus enhancing the differentiation of stem cells towards the osteogenic phenotype.However,the effects of macropore size on osteogenic differentiation of stem cells under dynamic condition are still unclear.Therefore,the objective of this study was to investigate the effects of macropore size of hydroxyapatite(HAp)scaffolds on osteogenic differentiation of bone mesenchymal stem cells under static and perfusion culture conditions.In vitro cell culture results showed that cell proliferation,alkaline phosphate(ALP)activity,mRNA expression of ALP,collagen-I(Col-I),osteocalcin(OCN)and osteopontin(OPN)were enhanced when cultured under perfusion condition in comparison to static culture.Under perfusion culture condition,the ALP activity and the gene expression of ALP,Col-I,OCN and OPN were enhanced with the macropore size decreasing from 1300 to 800 mm.However,with the further decrease in macropore size from 800 to 500 mm,the osteogenic related gene expression and protein secretion were reduced.Computational fluid dynamics analysis showed that the distribution areas of medium-and high-speed flow increased with the decrease in macropore size,accompanied by the increase of the fluid shear stress within the scaffolds.These results confirm the effects of macropore size on fluid flow stimuli and cell differentiation,and also help optimize the macropore size of HAp scaffolds for bone tissue engineering.展开更多
Hybridoma cells were cultured by continuous perfusion in Fibra-Cel of 5L packed-bed bioreactor for 22 days in low serum or serum-free media.The corresponded amino acids were fed and serum concentration was decreased b...Hybridoma cells were cultured by continuous perfusion in Fibra-Cel of 5L packed-bed bioreactor for 22 days in low serum or serum-free media.The corresponded amino acids were fed and serum concentration was decreased by analyzing glucose concentration, oxygen uptake rate, secretary antibody amount and amino acids concentration in culture supernatant. Comparing with continuous perfusion culture that amino acids were not fed, antibody amunt of production was increased about 2～3 times. The inoculated cell density was 2.5×10 5 cells/mL,while the final cell density was 8.79×10 8cells/mL. Antibody production was reached 295mg/L/d at average level, and the highest level was reached 532mg/L/d.These results provided a primary mode of enlarge culture for monoclonal antibody industrilization.展开更多
基金Supported by The European Union Grant STREP NMP3-CT-29005-013811(to Welle A)the Bundesministerium für Bildung und Forschung Grant 03ZIK-465(to Altmann B),Germany
文摘One of the greatest impacts on in vitro cell biology was the introduction of three-dimensional(3D)culture systems more than six decades ago and this era may be called the dawn of 3D-tissue culture.Although the advantages were obvious,this field of research was a "sleeping beauty"until the 1970s when multicellular spheroids were discovered as ideal tumor models.With this rebirth,organotypical culture systems became valu-able tools and this trend continues to increase.While in the beginning,simple approaches,such as aggregation culture techniques,were favored due to their simplicity and convenience,now more sophisticated systems are used and are still being developed.One of the boosts in the development of new culture techniques arises from elaborate manufacturing and surface modification tech-niques,especially micro and nano system technologies that have either improved dramatically or have evolved very recently.With the help of these tools,it will soon be possible to generate even more sophisticated and more organotypic-like culture systems.Since 3D per-fused or superfused systems are much more complex to set up and maintain compared to use of petri dishes and culture flasks,the added value of 3D approaches still needs to be demonstrated.
基金supported by National Natural Science Foundation of China(82002289)the Medical Research Project Plan of Sichuan Province(S20012)+4 种基金Sichuan Science and Technology Program(21MZGC0218)Sichuan Science and Technology Innovation Seeding Project(2020065)the College-City Cooperation Project of Nanchong City(19SXHZ0236,19SXHZ0099,20SXQT0335)Startup Program of China West Normal University(18Q069,18Q030)National Natural Science Foundation of North Sichuan Medical College(CBY19-YZ09).
文摘Previous studies have proved that dynamic culture could facilitate nutrients transport and apply mechanical stimulation to the cells within three-dimensional scaffolds,thus enhancing the differentiation of stem cells towards the osteogenic phenotype.However,the effects of macropore size on osteogenic differentiation of stem cells under dynamic condition are still unclear.Therefore,the objective of this study was to investigate the effects of macropore size of hydroxyapatite(HAp)scaffolds on osteogenic differentiation of bone mesenchymal stem cells under static and perfusion culture conditions.In vitro cell culture results showed that cell proliferation,alkaline phosphate(ALP)activity,mRNA expression of ALP,collagen-I(Col-I),osteocalcin(OCN)and osteopontin(OPN)were enhanced when cultured under perfusion condition in comparison to static culture.Under perfusion culture condition,the ALP activity and the gene expression of ALP,Col-I,OCN and OPN were enhanced with the macropore size decreasing from 1300 to 800 mm.However,with the further decrease in macropore size from 800 to 500 mm,the osteogenic related gene expression and protein secretion were reduced.Computational fluid dynamics analysis showed that the distribution areas of medium-and high-speed flow increased with the decrease in macropore size,accompanied by the increase of the fluid shear stress within the scaffolds.These results confirm the effects of macropore size on fluid flow stimuli and cell differentiation,and also help optimize the macropore size of HAp scaffolds for bone tissue engineering.
文摘Hybridoma cells were cultured by continuous perfusion in Fibra-Cel of 5L packed-bed bioreactor for 22 days in low serum or serum-free media.The corresponded amino acids were fed and serum concentration was decreased by analyzing glucose concentration, oxygen uptake rate, secretary antibody amount and amino acids concentration in culture supernatant. Comparing with continuous perfusion culture that amino acids were not fed, antibody amunt of production was increased about 2～3 times. The inoculated cell density was 2.5×10 5 cells/mL,while the final cell density was 8.79×10 8cells/mL. Antibody production was reached 295mg/L/d at average level, and the highest level was reached 532mg/L/d.These results provided a primary mode of enlarge culture for monoclonal antibody industrilization.
