ORIGINAL ARTICLE

Caco-2 cells monolayer as an in-vitro model for probiotic strain translocation

Ni Nengah Dwi Fatmawati, Kazuyoshi Goto, I Putu Bayu Mayura, Komang Ayu Nocianitri, Yan Ramona, Masakiyo Sakaguchi, Osamu Matsushita, I Nengah Sujaya

Ni Nengah Dwi Fatmawati
Department of Microbiology, Faculty of Medicine, Universitas Udayana, Bali, Indonesia

Kazuyoshi Goto
Department of Bacteriology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Japan

I Putu Bayu Mayura
Department of Microbiology, Faculty of Medicine, Universitas Udayana, Bali, Indonesia Department of Bacteriology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Japan

Komang Ayu Nocianitri
Department of Food Science Technology, Faculty of Agricultural Technology, Universitas Udayana, Bali, Indonesia Integrated Laboratory Biosciences and Biotechnology, Universitas Udayana, Bali, Indonesia

Yan Ramona
Integrated Laboratory Biosciences and Biotechnology, Universitas Udayana, Bali, Indonesia School of Biology, Faculty of Mathematics and Sciences, Universitas Udayana, Bali, Indonesia

Masakiyo Sakaguchi
Department of Cell Biology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Japan

Osamu Matsushita
Department of Bacteriology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Japan

I Nengah Sujaya
Integrated Laboratory Biosciences and Biotechnology, Universitas Udayana, Bali, Indonesia School of Public Health, Faculty of Medicine, Universitas Udayana, Bali, Indonesia. Email: nsujaya@unud.ac.id
Online First: April 01, 2020 | Cite this Article
Fatmawati, N., Goto, K., Mayura, I., Nocianitri, K., Ramona, Y., Sakaguchi, M., Matsushita, O., Sujaya, I. 2020. Caco-2 cells monolayer as an in-vitro model for probiotic strain translocation. Bali Medical Journal 9(1): 137-142. DOI:10.15562/bmj.v9i1.1633


Background: Caco-2 cells monolayer is one of invitro models to evaluate the translocation capacity of Lactobacillus spp probioticstrains. The translocation is influenced by mucosa permeability of enterocytes as shown by increasing transepithelial resistance (TER) and formation of tight junction proteins. The pore size of the supported permeable membrane used in in vitro assay was one of the crucial factors in performing bacterial translocation assay. Almost no study has been conducted using Caco-2 cells monolayer grown on 8-μm pore size polycarbonate membrane for evaluating probiotics translocation. Therefore this study aimed to determine whether the Caco-2 cells monolayer model was suitable as an in vitro translocation model.

Methods: Caco-2 cells monolayer was seeded onto 8-μm collagen-coated polycarbonate membrane insert Transwell®. Differentiation of Caco-2 cells was detected by measuring the TER, while the ZO-1 protein (the tight junction proteins) was detected by immunofluorescence. H2O2 was used as a tight junction disruptive agent. Data were analyzed using SPSS version 23 software to compare the mean of TER measurement between untreated and H2O2-treated Caco-2 cells monolayer.

Results: The result showed that the TER of Caco-2 cells monolayer was gradually increasing until day 14, reaching more than 800 ohm.cm2. Furthermore, the ZO-1 protein was successfully detected, indicated the tight junction formation.  TER value of H2O2-treated cells showed significantly lower than that of untreated cells (P<0.05), indicating a disturbance of cells monolayer integrity. Lactobacillus rhamnosus FBB81 was used for validating the translocation. There was no translocation observed; however, translocation was observed in H2O2-treated cells.

Conclusion: Altogether suggests that Caco-2 cells grown on 8 μm-pore size permeable filters could be considered as a suitable in vitro model for probiotics strains translocation.

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