Groove shape porous plate animal cell hollow fiber reactor and use thereof
An animal cell, multi-well plate technology, applied in bioreactor/fermenter combination, specific-purpose bioreactor/fermenter, tissue cell/virus culture device, etc., can solve problems such as inability to meet, and achieve simplified operation Steps, effects of good toxicity or protection
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specific Embodiment 1
[0025] Specific embodiment 1: (comparison on isoniazid drug toxicity)
[0026] Rat hepatocytes with a live rate higher than 90% were harvested by 10 6 The cells / ml density was inoculated in a mixture of 4-fold concentrated medium and rat tail collagen solution (1:3; v:v), injected into hollow fiber 3, and placed at 37°C for 10 minutes to solidify. Cut the hollow fiber 3 short, 6.5cm × 12, and put each 3 into the groove of a 24-slot orifice plate, add 3ml of Williams'E medium containing 5% fetal bovine serum, put it into 37 ° C, 5% CO 2 Culture in an incubator.
[0027] The same batch of rat liver cells was divided into 0.8×10 6 Cells / ml were inserted into 24-well cell culture plate with mouse tail collagen in 3ml Williams'E medium containing 5% fetal bovine serum. The culture conditions were the same as above.
[0028] After the cells adhered to the wall in the 24-well cell culture plate (12-18h), the medium containing the drug to be tested isoniazid and the medium withou...
specific Embodiment 2
[0031] Specific embodiment 2: (for the evaluation of hepatoprotective drug)
[0032] Rat hepatocytes with a live rate higher than 90% were harvested by 10 6 The cells / ml density was inoculated in a mixture of 4-fold concentrated medium and rat tail collagen solution (1:3; v:v), injected into hollow fiber 3, and placed at 37°C for 10 minutes to solidify. Cut the hollow fiber 3 short, 6.5cm × 12, and put each 3 into the groove of a 12-slot slotted orifice plate, add 3ml of Williams'E medium containing 5% fetal bovine serum, put it into 37 ° C, 5% CO 2 Culture in an incubator.
[0033] After culturing for 12-18 hours, each group was replaced with medium containing 0.25mM tacrine, 0.25mM tacrine+0.5g / l glycyrrhizic acid, and no drug (as a blank control). Observing the cell morphology after 48 hours, it was found that the cell viability and glutathione of 0.25mM tacrine + 0.5g / l glycyrrhizic acid were higher, close to the blank control, and compared with the tacrine group, there...
specific Embodiment 3
[0034] Specific embodiment 3: (for drug metabolism research)
[0035] Human hepatocytes with a viability higher than 90% were harvested by 10 6 The cells / ml density was inoculated in a mixture of 4-fold concentrated medium and rat tail collagen solution (1:3; v:v), injected into hollow fiber 3, and placed at 37°C for 10 minutes to solidify. Cut the hollow fiber 3 short, 6.5cm × 12, and put each 3 into the groove of a 24-well slotted orifice plate, add 3ml of Williams'E medium containing 5% fetal bovine serum, put it into 37 ° C, 5% CO 2 Culture in an incubator.
[0036] The same batch of rat liver cells was divided into 0.8×10 6 cells / ml were inserted into the culture wells of 2ml of Williams'E medium containing 5% fetal bovine serum and 24-well cell culture plate covered with rat tail collagen. The culture conditions were the same as above.
[0037] After culturing for 60 h, each group was added drug metabolism substrates (40uM phenacetin solution and 4-nitrocatechol, 4-...
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