Projects / Programmes
Use of Adipose-derived Stem Cells for Engineering Vascularized Tissue Implants
| Code |
Science |
Field |
Subfield |
| 4.06.02 |
Biotechnical sciences |
Biotechnology |
Bio-engineering |
| Code |
Science |
Field |
| B000 |
Biomedical sciences |
|
| Code |
Science |
Field |
| 3.04 |
Medical and Health Sciences |
Medical biotechnology |
stem cells, endothelial cells, adipose tissue, stromal vascular fraction, vascularization, tissue implants, cell therapies, tissue engineering
Researchers (1)
| no. |
Code |
Name and surname |
Research area |
Role |
Period |
No. of publicationsNo. of publications |
| 1. |
25484 |
PhD Mirjam Fröhlich |
Biotechnology |
Head |
2012 - 2015 |
55 |
Organisations (2)
Abstract
Numerous tissue defects, which arise due to diseases or injuries, represent one of the major clinical problems. The need for tissues will be in coming years even increasing, due to the aging of human population. Tissue engineering (TE) approach could provide autologous tissue implants for the transplantation, thus alleviating the shortage of tissues. In TE approach, cells are harvested from a small biopsy of the patient, are subsequently expanded in vitro, combined with a suitable biomaterial (scaffold) and are finally implanted back to the patient, to the site of the injury. Some clinical cases are already reporting good results using TE to treat various tissue defects, yet, many issues still need to be addressed. One of most important problems to be solved if TE approach is to be used in clinical practice is the vascularization. Several studies prove that implantation of tissue implant fail, because the implant doesn’t integrate with the surrounding tissue. The main reason for this is the lack of the vascular network within the implant. The vasculature assures the exchange of nutrients and gases and is therefore of crucial importance for the survival of the tissue implants after they have been implanted. Besides its nutritional role, the vasculature is also of great importance for the correct maturing of other cell types within the tissue. Some examples of the tissues which require extensive vascular network for their functioning are bone, adipose, and muscle tissue.
Endothelial cells for the purpose of vascularization could be obtained from different sources, such as umbilical vein or from peripheral blood. Since both cell sources have important clinical drawbacks (in first case biological unsuitability, and in second limited quantity), the alternative source of endothelial cells is needed. In the proposed study we would like to overcome disadvantages of current TE approaches, such as lack of vasculature and less suitable cell sources. The overall aim of the study is to evaluate the vascular potential of cells from adipose tissue (endothelial cells, stem cells, pericytes) as a cell source for the purposes of vascularization. Quantity of cells is often a critical parameter, therefore the great advantage of adipose tissue is its quantity and the ease of access. We are aiming for preparing of a platform for vascularization of (i) bone tissue, (ii) subcutaneous/adipose tissue, and (iii) heart muscle. Principal methods employed in the study will be: isolation of cells from tissues and establishment of in vitro cell cultures under various conditions (hypoxia, perfusion bioreactors), microscopy (light, fluorescence), immunocytochemical techniques, molecular (qPCR) and biochemical (ELISA) methods. Specific goals of the project are the following: (i) to define the most optimal anatomical site (tummy, buttocks, thighs) for the harvesting of the largest possible quantities of functional endothelial cells, (ii) to define in vitro protocols for adipose derived endothelial cell proliferation, (iii) to define the portion of endothelial cells in SVF and the portion of endothelial cells which differentiate from stem cells (ASC), (iv) to assess the effects of hypoxia on proliferation and differentiation capabilities of endothelial cells, (v) to evaluate interactions of endothelial cells with other tissue specific cell types (bone -, smooth muscle -, adipose cells), and finally (vi) to introduce the proposed vascularization approach as a novel method to the clinical practice.
Successfully implemented study will importantly contribute to the development of the area of cell therapies and tissue engineering. The potential introduction of the innovative approaches into clinical practice will result in socio-economical benefits for Slovenian, as well as for international society.
Significance for science
Due to numerous tissue defects, which will even increase further in coming years due to the aging of the population, there is a great need for tissue substitutes. Bone defects, and soft tissue defects present two big areas for the use of in vitro engineered tissue implants. The success of implantation of the in vitro engineered tissue greatly depends on the vascularization of the implant. The problem of vascularization, however, hasn’t been adequately solved yet. In our studies, all cell types needed for engineering a tissue implant can be obtained from a single, clinically relevant cell source – adipose tissue, which is easy to harvest and is safe for the patient. The focus of our research were the endothelial cells derived from stromal vascular fraction, namely, their characteristics (quantity, origin, self-assembly potential to form vascular structures), their response to specific environments characteristic for the implantation site or the process of preparation of cell product (such as hypoxia, presence or absence of VEGF, osmolarity), and their integrations with other tissue specific cells. The obtained results contribute to the development of scientific field of stem cells, endothelial cells, and cell interactions, as well as to the area of in vitro vascularization approaches, which are applicable to the broad field of cell therapy and tissue engineering aiming at regeneration of bone, soft (skin) tissues, heart and skeletal muscle, vessels, wounds, etc. Results of this study also served as a base for the development of new cell technology platform aiming for regeneration of bone and augmentation of soft tissue.
Significance for the country
The results of the study serve as a basis for the development and introduction of new innovative approaches for the regeneration of tissues into the clinical practice. The novel approach will contribute to solving one of the most difficult problems in the field of cell and tissue engineering – vascularization. The results and knowledge gained through the project are applicable to the broad field of cell therapy and tissue engineering aiming at regeneration of bone, soft (skin) tissues, heart and skeletal muscle, vessels, wounds, etc. The main Educell’s activity is development of cell therapies for regeneration of tissues in humans. In this context, based on the project results, a new cell technology platform aiming at regeneration of bone and soft tissue has been developed by the company and was successfully introduced into the clinics already in the course of the project. Furthermore, results present the possibility of development of additional cell products, as well as quality control tests and therefore contribute to direct professional and economical benefits of the company, as well as growth and promotion of whole economy branch – biotechnology, more specifically tissue engineering. Introduction of more efficient methods to the clinical practice also has a beneficial socio-economical impact as they result in fewer secondary interventions needed, which is consequently leading to money savings, as well as to improving the life quality of the patients. In the course of the project, numerous collaborations were initiated or maintained, including world-wide acknowledged research laboratories and industry, which contributed to the increased quality of results and increase of the competitiveness of Slovenian science as well as the business sector. Successful implementation of the project and introduction of new methods to the clinical practice importantly promotes the company, research entities, the country and its scientists internationally, as examples of successful implementation of cell therapies in the clinics are still fairly rare. New knowledge and development of tissue engineering approaches contributes to the development of tissue engineering – currently, a rather undeveloped field in Slovenia - which might provide additional chances for the education and employment of younger people. The knowledge will also be transferred through lectures to the students of biotechnology at the Biotechnical Faculty of the University of Ljubljana.
Most important scientific results
Annual report
2012,
2013,
final report,
complete report on dLib.si
Most important socioeconomically and culturally relevant results
Annual report
2012,
2013,
final report,
complete report on dLib.si
