title: Optimization of Caco-2 and HT29 co-culture in vitro cell models for permeability studies.
abstract: The purpose of this study was to investigate the appropriate proportion of Caco-2 and HT29 co-culture in vitro cell models for permeability studies. The results showed that the transepithelial electrical resistance values of 9:1 and 1:0 groups (263 ± 3.61 and 300 ± 7.55) after 21-day culture were >250 Ω cm(2), which were suitable for further experiments. The confocal laser microscopy showed that the group of 9:1 (Caco-2:HT29) had the highest integrity, whereas the group of 0:1 (Caco-2:HT29) exhibited the lowest. The staining study confirmed that mucus was successfully produced by HT29 cells, and it was also produced in co-cultures with Caco-2 cells model, but the Caco-2 monocultures did not have any blue staining, which made us affirm that mucus is only produced in the presence of HT29 cells. The real-time PCR results showed that the total highest expression level of ALPi and MUC5AC was the ratio of 9:1 (Caco-2:HT29) and lowest is 1:1 (Caco-2:HT29). So we concluded that 9:1 (Caco-2:HT29) is the optimal Caco-2 to HT29 ratio in the in vitro model co-culture for permeability studies.
authors: a Laboratory of Nutrition and Functional Food , Jilin University , Changchun , PR China.
authors: Fengguang
authors: F
authors: Pan
authors: a Laboratory of Nutrition and Functional Food , Jilin University , Changchun , PR China.
authors: Lu
authors: L
authors: Han
authors: a Laboratory of Nutrition and Functional Food , Jilin University , Changchun , PR China.
authors: Yan
authors: Y
authors: Zhang
authors: a Laboratory of Nutrition and Functional Food , Jilin University , Changchun , PR China.
authors: Yiding
authors: Y
authors: Yu
authors: a Laboratory of Nutrition and Functional Food , Jilin University , Changchun , PR China.
authors: Jingbo
authors: J
authors: Liu
mesh_terms: D000423:Alcian Blue; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism*; D018938:Caco-2 Cells; D018920:Coculture Techniques; D004396:Coloring Agents; D017097:Electric Impedance; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D019073:HT29 Cells; D006801:Humans; D055271:Mucin 5AC / Q000235:genetics / Q000378:metabolism*; D009093:Mucus / Q000378:metabolism*; D010539:Permeability; D060888:Real-Time Polymerase Chain Reaction; D013194:Staining and Labeling
chemical_list: D004396:Coloring Agents; D058851:GPI-Linked Proteins; C524678:MUC5AC protein, human; D055271:Mucin 5AC; C063088:ALPI protein, human; D000469:Alkaline Phosphatase; D000423:Alcian Blue
keywords: Cell seeding ratio; co-culture; gene expression; monolayer; optimization