Projects / Programmes
Induced pluripotent stem cells in goat.
| Code |
Science |
Field |
Subfield |
| 4.02.01 |
Biotechnical sciences |
Animal production |
Genetics and selection |
| Code |
Science |
Field |
| B400 |
Biomedical sciences |
Zootechny, animal husbandry, breeding |
| Code |
Science |
Field |
| 4.02 |
Agricultural and Veterinary Sciences |
Animal and Dairy science |
reprogamming, induced pluripotent stem cells, pluripotency, mammary gland
Researchers (1)
| no. |
Code |
Name and surname |
Research area |
Role |
Period |
No. of publicationsNo. of publications |
| 1. |
28505 |
PhD Jernej Ogorevc |
Animal production |
Head |
2013 - 2015 |
137 |
Organisations (1)
Abstract
BACKGROUND: Induced pluripotent stem cells (iPSCs) are stem cells obtained by direct reprogamming of differentiated (somatic) cells. iPSCs show similarities to embryonic stem cells (ESCs), which are in farm animals very difficult to isolate and maintain in cell culture, due to the lack of specific knowledge. Because of it's short gestation period and early sexual maturation goat is an excellent model for studying fetal development and lactation and of great importance for biopharming. Cell types present in goat mammary gland, which undergo cyclic changes during repeated lactations and involutions (differentiation, proliferation, and apoptosis) seem appropriate model to study reprogamming. Furthermore, mammary cells were already proven to have reprogamming potential (cloning of Dolly), while little is known about direct reprogamming of these cells. PROBLEM: Until recently it was possible to obtain stem cells only from embryos in the early stages of development (ESCs), which causes ethical concerns and generates legal restriction. iPSCs are derived from somatic cells, therefore, are ethically acceptable and represent almost unlimited source of pluripotent cells. The process of reprogamming allows production of stem cells which are genetically identical to individuum. Before the potential of iPSCs could be used in medicine it is necessary to explain genetic and epigenetic regulation of reprogamming. Due to morphological and physiological similarities to human farm animals represent an excellent model for preclinical research. Additionally, there are no established ES cell lines in farm animals. iPSCs could serve as a substitute for ESCs to generate transgenic animals, which could be used for production of recombinant proteins in their mammary glands. AIMS: The project aims to explore the possibility of mammary epithelial cell type reprogamming, with the aim to determine which exogenous transcription factors are required for efficient cell reprogamming, to confirm pluripotent character of reprogammed cells, and to assess their tumorigenic potential. Since it appears that reprogamming process is conserved across species, we will compare the obtained results with results obtained in other species and try to contribute to the understanding of reprogramming. METHODS: From the data, previously obtained by sequencing the entire transcriptome of goat mammary epithelial cells, information about potential endogenous expression of pluripotency transcription factors in this particular cell type will be extracted. This will aid in determining which exogenous transcription factors needs to be inserted. Pluripotent potential of reprogammed cells will be verified by genetic, epigenetic and functional tests. SIGNIFICANCE: Pluripotent cells are particularly important for regenerative medicine, for drug development and testing, for basic research, and for conservation of endangered species. In species, where ESCs are hard to isolate and maintain, iPSCs could be used for generation of transgenic animals. However, before it will be possible to exploit their full potential reprogamming processes should be explained and safe methods developed, that will enable the production of genetically and epigenetically stable cells. ORGANIZATION AND FEASIBILITY: Existing data on the transcriptome profile of goat mammary epithelial cells, candidates experience in the field of mammary gland biology and stem cells, and the availability of well-equipped laboratories enables good organization and feasible realization of the project.
Significance for science
In the last years significant progress has been made in the field of stem cell research and reprogramming strategies. Reprogramming technology allows generation of pluripotent individuum-specific stem cells in unlimited amounts that is not dependent on destruction of embryos. Currently, effective methods that allow generation of pluripotent cells with germline competence, are available only for rodents. Our experiment was the first try of direct reprogramming of mammary epithelial cells and one of the first in goats. Further development of technology that would allow use of reprogrammed cells in human medicine, requires preclinical testing in large farm animals, which are more related to human than murines. Research on large farm animals will facilitate the development of new reprogramming strategies and derivation and characterization of iPSCs in different species. iPSCs from different species will eventually reveal optimal culture conditions, safe reprogamming methods and suitable markers for generation of pluripotent cells in different species. Additionally, iPSCs in goats, where no ESCs lines are available, could be used as an ESC alternative for production of transgenic goats. Animal pluripotent stem cells can be genetically modified in in vitro conditions and used in combination with advanced reproduction techniques (e.g. SCNT, blastocyst complementation) for production of animals with special traits. Transgenic animals could be used in agriculture or as bioreactors for production of recombinant proteins. Ruminants (especially goats) are important species for biopharming, because expression of transgenic proteins in mammary gland is currently the most optimal production system that allows recombinant proteins to be relatively easy isolated from milk. Results of the project and rare literature reports show that reprogramming of goat (ruminant) cells seems more difficult as reprogramming of human or mouse fibroblasts, and probably requires insertion of additional reprogramming factors and optimization of reprogramming medium. In the course of this project an important step forward was achieved in characterisation of ruminant mammary cells and towards their successful reprograming.
Significance for the country
Reprogramming somatic cells back to pluripotency is one of the most important research topics in modern science and represents a great potential for further research in basic science and applicative uses in agriculture, pharmacy, and medicine. Field of cell reprogramming hasn't been subjected to a significant amount of research in Slovenia, therefore it is necessary to increase international competitiveness of researchers in this field. Accessibility of pluripotent stem cells in animals can aid in deciphering and understanding reprogramming mechanisms and enables preclinical research on animal models. Additionally, goat pluripotent stem cells can be used for development of transgenic goats that can be of high value as bioreactors in biopharmaceutical industry. Findings of this project are a starting point for further research and possible new projects in the field.
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