Lab-scale Production of Cell-free Probiotic Supernatant Services

Creative Biolabs' lab-scale custom production services are flexible, reliable, and easy to use to facilitate research projects. Our scientists have extensive experience and excellent skills in the field of probiotics. Our goal is to provide our customers with affordable service and reliable results.


Gut microbiota plays a dual role in protecting host health by producing a variety of metabolites and biologics. Probiotics are known to produce many metabolites with antibacterial and antibiofilm activity that are often secreted into the surrounding media, such as bacteriocins and organic, antimicrobial peptides and fatty acids, biosurfactants, and hydrogen peroxide. These metabolites are often collected from the cell-free supernatants, and thus the mechanism of action of supernatants is directly related to the antimicrobial metabolites produced by probiotics, which may differ among species. Dead probiotics and their metabolites also have significant benefits in the prevention and treatment of tumor cells. To determine whether probiotics can prevent or treat a specific pathogen infection, two types of studies can be conducted: prophylactic studies (clinical studies that examine whether exposure to the pathogen has been prevented after the use of the probiotics) or interventional studies (prior exposure to the pathogen followed by treatment with the probiotics or their supernatant).

Role of Cell-free Supernatants

The supernatants produced by different microbial cultures have different activities. Supernatants derived from bacterial cultures of the genera Lactobacillus and Bifidobacterium were also recently shown to display antibacterial activity in vitro. Therefore, cell-free bacterial supernatants are promising anti-infective agents for, for example, the treatment of diarrhea.

  • Anti-C. diffificile

It has been proved that cell-free supernatants obtained from Lactobacillus delbrueckii ssp. bulgaricus B-30892 and different Bifidobacterial strains were able to reduce the adhesion of C. difficile to intestinal epithelial Caco-2 cells.

  • Anti-colitis

A study showed decreased IL-8 concentrations and the abolishment of NFκB activation in Caco2 cells when exposed to the supernatant fraction of Faecalibacterium prausnitzii. Moreover, the researchers demonstrated that the cell-free supernatant added to a colitis mouse model could shift the immune response toward an anti-inflammatory pattern characterized by an increase in IL-10 and a decrease in IL-12.

  • Anti-cancer

Molecules and metabolites derived from probiotics can prevent tumor development through the modulation of the immune systems of the host. B. bifidum metabolites, cell-free supernatants are fighting tumor cells effectively when using colon cancer cell line SW742. Cell-free supernatants (CFS) from L. casei and L. rhamnosus GG have been shown to prevent colon cancer cell invasion suggesting that probiotic CFS has anti-metastatic bioactive substances that may participate in cell invasion decrease in vitro.

  • Against Pathogenic Biofilm

The scientist demonstrated that the pH non-neutralized culture supernatant of isolates of Lactobacillus spp. could prevent the biofilm formation of V. cholera. In addition, in the presence of Lactobacillus fermentum supernatant, a thin layer of S. aureus biofilm was formed on the glass surface instead of the thicker layer of the control. The supernatant's ability to prevent biofilm formation appears to be higher than its ability to destroy preformed biofilm.

Schematic representation of various health benefits of postbiotic molecules.Fig.1 Schematic representation of various health benefits of postbiotic molecules. (Nataraj, 2020)

Creative Biolabs' Services for Probiotic Cell-free Supernatants Preparation

Cell-free supernatants containing biologically active metabolites secreted by bacteria and yeast into the surrounding liquid can be obtained from cell cultures. Creative Biolabs can grow probiotics according to customers' requirements, obtain probiotic cell-free supernatant, and study the effects of probiotic metabolites or animal research. Including but not limited to the following probiotics:


  • Fully customized
  • Flexible
  • Fast delivery
  • Experienced

With Creative Biolab as your CRO partner, we can create valuable insights through strong scientific knowledge of microbiology, probiotics, live biotherapeutics, and an advanced technology platform. Find out how we can help you achieve your goals.


  1. Nataraj, B.H.; et al. Postbiotics-parabiotics: The new horizons in microbial biotherapy and functional foods. Microbial cell factories. 2020, 19(1): 1-22.

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