Stem Cells

Research title: 

The stem cell niche in domestic animal intestinal mucosa: biology and applications. 

Tutor: Prof. Fulvio Gandolfi

 

Contact details

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Via Celoria, 2 - 20133 Milano (Italy)

University of Milan - Department of Agricultural and Environmental Sciences - Production, Landscape, Agroenergy.  http://eng.disaa.unimi.it/ecm/home

Tel. +39 02 5031 7990

Fax +39 02 5031 7980

 

State of the art

A clear relationship has been established between integrity and functionality of the gastrointestinal wall and feed intake. Lower feed leads to reduced body weight gain and an increased likelihood for opportunistic pathogens to attack the digestive tract and cause diarrhea [1].

Antibiotics used to be included in animal diets to prevent pathologies and optimize growth performance especially in intensive farming. However, owing to the possible contribution of in-feed antibiotics to the development of antibiotic-resistant strains of bacteria, the European Union implemented a full ban on in-feed antibiotics usage in livestock diets in 2006.

Therefore, it is important to find alternatives/replacements to in-feed antibiotics that are effective in reducing the incidence and severity of digestive problems. The intestinal tract complexity, makes it difficult to forecast the efficacy of a substance or a combination without extensive in vivo trials. Restrictive rules and very high costs limit their use and the number of substances that can be tested for efficacy and safety. The development of in vitro tests able to efficiently screen compounds selected on the basis of theoretical considerations is desirable.

To this purpose, a better knowledge of the inner epithelial lining of the small intestine is required together with the development of sophisticated 3-D culture systems that can closely mimic the intestinal mucosa.

Recently, robust adult intestinal stem cell markers, enabled an improved understanding how intestinal stem cells interact with their local niche to maintain homeostasis in healthy intestinal epithelia, in mouse and in humans whereas a limited amount of information are available in domestic species.

 

gandolfi1

 

Figure 1. Pig Intestinal mucosa in the duodenum (A) and immunolocalization of villin in the same region (B).

gandolfi2

Figure 2. Scheme of an intestinal organoid and two examples of pig duodenum and jejunum crypto-villi structures in culture

 

Aims of the project

The currently available immortal cell lines made of a single cell type poorly represent of the intestinal mucosa complexity. As an alternative, in recent years’, intestinal organoids have been developed in mouse and humans. These structures are formed by different cell types and include the intestinal stem cell niche that has recently been characterized in vivo. These are currently considered the most advanced in vitro experimental model. At present, however, such detailed and extended knowledge is not available in the domestic species. A few preliminary results have been recently described by Gonzales and coll. [2] who validated a number of cell-type specific markers for the pig. These data were obtained exclusively from 6-8 week-old individuals kept in experimental facilities. Therefore, representing a narrow window of the pig life span and from animals which were maintained is conditions that do not reflect the normal procedures of modern animal production.

Therefore aims of this project are:

  • to characterize the stem cell niche in domestic species;
  • to develop intestinal organoids in these species;
  • to determine the ability of these organoids to provide relevant information for the screening of candidate compounds able to replace in-feed antibiotics.

We will initially work on pigs but we will consider the opportunity to extend our research to poultry and farmed fishes.

 

[1] Feoli C. Early nursery nutrition – getting pigs started. London Swine Conf. – A Platf. Success April 5 April 6, 2016, London, Ontario CA: 2016, p. 145–50.

[2] Gonzalez LM, Williamson I, Piedrahita JA, Blikslager AT, Magness ST. Cell lineage identification and stem cell culture in a porcine model for the study of intestinal epithelial regeneration. PLoS One 2013;8:e66465. doi:10.1371/journal.pone.0066465.

 

Recent publications of the tutor in the field

1) Brevini, T.A., Pennarossa, G., Acocella, F., Brizzola, S., Zenobi, A., Gandolfi, F., 2016. Epigenetic conversion of adult dog skin fibroblasts into insulin-secreting cells. Vet J 211, 52-56.

2) Maffei, S., Galeati, G., Pennarossa, G., Brevini, T.A., Gandolfi, F., 2015. Extended ex vivo culture of fresh and cryopreserved whole sheep ovaries. Reprod Fertil Dev DOI 10.1071/RD15101.

3) Manzoni, E.F., Pennarossa, G., deEguileor, M., Tettamanti, G., Gandolfi, F., Brevini, T.A., 2016. 5-azacytidine affects TET2 and histone transcription and reshapes morphology of human skin fibroblasts. Sci Rep 6, 37017.

4) Pennarossa, G., Maffei, S., Campagnol, M., Rahman, M.M., Brevini, T.A., Gandolfi, F., 2014. Reprogramming of Pig Dermal Fibroblast into Insulin Secreting Cells by a Brief Exposure to 5-aza-cytidine. Stem Cell Rev 10, 31-43.

5) Pennarossa, G., Zenobi, A., Gandolfi, C.E., Manzoni, E.F., Gandolfi, F., Brevini, T.A., 2016. Erase and Rewind: Epigenetic Conversion of Cell Fate. Stem Cell Rev 12, 163-170.