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Keywords
EMBRYONIC STEM CELLS, PRIMORDIAL STEM CELLS, ADULT STEM CELLS, IN VITRO DIFFERENTIATION, IN VIVO DIFFERENTIATION, NUCLEAR TRANSPLANTATION, PIG, SHEEP, SCID MICE

Cell therapy: development of biomedical and pre-clinical models using non laboratory animals

Università degli Studi di Milano
Abstract
A stem cell can both self-renew, in an undifferentiated state, and differentiate into one or more cell types. Stem cells are defined as totipotent or pluripotent when can originate all or most cell types that compose an organism, as is the case in the early embryo. Stem cells exist also in the adult, these cells however can give rise to a limited number of cell types and are defined as multipotent.
Stem cells hold a great potential both for research and clinical applications. Whereas studies on basic mechanisms are better performed with mouse cells, clinical applications would greatly benefit by the development of alternative animal models to be used beside the mouse which could provide information more relevant for the human species.
The use of traditional laboratory animals (mouse, rat, rabbit) needs to be followed by pre-clinical trials based on experimental models whose morphology, physiology and genetic background are more similar to humans. Pig and sheep have been used for many years as suitable models in different branches of medicine. Therefore it is necessary to develop these models also in the field of stem cell therapy deriving stem cells of different origins in these species.

Aim of this project is the derivation and characterization of pig and sheep stem cell lines in order to use them in pre-clinical trial for the development of cell therapy for human and veterinary applications.

The four units involved in the project >>>

Principal Investigator
Fulvio Gandolfi Università degli Studi di MILANO
Research Objectives
Aim of the present project is to derive pig and sheep stem cell lines of different origin in order to progress toward the clinical application of cell therapy in human and veterinary medicine.

There different kind of stem cells will be derived and characterized:
• pig and sheep embryonic stem cells
• sheep primordial germ cells
• sheep adult mesenchimal stem cells

For each cell line, aim of the project is:
• to improve the derivation procedures
• to evaluate the degree of self renewal through the analysis of specific markers
• to optimize the culture protocols for the establishment and maintenance of stable cell lines
• to identify the procedures for obtaining controlled cell differentiation in vitro
• to determine the degree of cell differentiation and integration following in vivo transplantation

Timescale
24 months
National and international background
A stem cell can both self-renew, in an undifferentiated state, and differentiate into one or more cell types [1]. Stem cells are defined as totipotent or pluripotent when can originate all or most cell types that compose an organism, as is the case in the early embryo. Stem cells exist also in the adult, these cells however can give rise to a limited number of cell types and are defined as multipotent [2].
Their possible application to cell therapy has recently generated great excitement [3]. Stem cells transplanted into a suffering organ have been shown to proliferate, give rise and replace the various damaged cell types and restore the original functions. Experiments carried out in laboratory animals have demonstrated stem cell ability to regenerate complex anatomical structures such as the brain [4] and the heart [5]. These observations have opened the way and given new expectations in the management of a number of lethal pathologies like Parkinson’s and Alzheimer’s diseases [6] or heart failure [7].

Stem cells can be of different origin

1. Embryonic Sten cells (ESC)

Embryonic stem cells are derived from embryos at the blastocyst stage, as originally shown in the mouse [8, 9] and recently demonstrated in human [10, 11]. Stem cells obtained in this way possess the highest possible differentiation plasticity and may be induced to differentiate into almost all cell types [12, 13]. Isolation of ES cells in species other than >>>