Cell-based In Vitro Assays for Live Biotherapeutics

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Essential Properties of Probiotics

The functional and specific positive biological effects of probiotics are strain-dependent. The most important properties of probiotics include acid and bile tolerance, adhesion to the human intestinal mucosa, temporary colonization of the human gastrointestinal tract, production of antimicrobial substances, and inhibition of the growth of pathogens. Probiotic bacteria's attachment to the intestine's wall and their colonization of the gastrointestinal tract is a vital precondition for their survival and functioning. Only probiotics with adhesive properties are thought to stabilize the intestinal mucosal barrier and contribute to the induction of immune effects. Adhesion may also administer routes of exclusion of harmful bacteria and keep them away from the epithelium.

Characterization and Screening of Candidate Probiotic

These screening models range from simple cell-based in vitro assays to complex ex vivo or animal models. In most cases, in vivo models cannot be used for high-throughput screening due to their high costs and due to ethical reasons. There is a wide range of in vitro assays being used to screen (candidate) probiotic strains. These in vitro assays were used because of their simplicity and relatively low cost. An important advantage of in vitro assays is their ease of use when multiple strains are screened simultaneously. Appropriate in vitro tests have been adopted to select strains, and in vitro assays can be used to study microbe-microbe interactions. Examples of in vitro assays are the use of human cell line assays to study bacterial adhesion. Cell lines like these have also been used to study the effects of microorganisms on epithelial barrier function. In probiotic research, in vitro co-culture assays are also frequently used to study interactions with the immune system.

Cell-based In Vitro Assays at Creative Biolabs

Cell culture models based on immortalized intestinal cell lines (Caco-2 or HT-29) can be used to evaluate electrical resistance and intestinal flux parameters. Transendothelial/transendothelial electrical resistance (TEER) is a widely accepted quantitative technique for measuring the integrity of tight junction dynamics in endothelial and epithelial monolayer cell culture models.

Investigating how probiotics influence cytokine networks and cytokine formation behaviors that are highly correlated with the initiation of adaptive immune responses is necessary to gain insight into the potentially beneficial effects of probiotics in promoting gut health.

The use of epithelial cell lines is very useful for the identification of adhesion mechanisms and molecules. In vitro cell line studies help predict the effect of probiotics and gastrointestinal conditions on the adhesion ability of probiotics.

Cell-based in vitro models for intestinal permeability studies.Fig.1 Cell-based in vitro models for intestinal permeability studies. (Pereira, 2016)

Creative Biolabs has always supported clients worldwide, from academic laboratories and small biotech start-ups to global biopharmaceutical companies. Whether you wish to commission a partner to provide you with full live biotherapeutics development or manage technical support for a single project, you can benefit from our expertise. Please do not hesitate to contact us for more details to support your research project.


  1. Pereira, C.; et al. Cell-based in vitro models for intestinal permeability studies. Concepts and models for drug permeability studies. 2016: 57-81.

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