This study investigates the changes in the pancreas of rats with the introduction of alloxan, confirming histologically the presence of pathological processes characteristic of type I diabetes. Additionally, a cytogenetic analysis of rat cells in culture obtained from red bone marrow, adipose tissue and pancreas during subcultivation in the in vitro system was performed and their phenotypic characterization was given. Furthermore the histological changes in the pancreas of rats were analysed with the introduction of cell cultures obtained from different sources against experimental diabetes and studied the changes in the blood glucose of experimental animals with the transplantation of cellular material.
Despite the shortcomings of the modern methods of treating animals with type I diabetes using insulin therapy, as well as the fact that pancreatic β-cell death is one of the important elements in the diabetes pathogenesis, new approaches to the treatment of this disease using cell technologies are being studied. Recent studies of β-cells in the in vitro system have shown that they have a fairly high regenerative capacity, but in the in vivo system with diabetes, these cells almost do not recover. The development of methods that can activate β-cell regeneration is an important area of the scientific research.
To date, mainly red bone marrow is used as the source of stem cells for research, because this is the only tissue of the adult body that normally contains immature, undifferentiated and low-differentiated cells. However, adipose tissue is being increasingly used as an alternative source for stem cells, from which they can be isolated in significantly larger quantities using less invasive methods compared to using red bone marrow. It is worth noting that there are still a lot of unclear issues in the study of the pancreas regeneration ways, therefore, in the treatment of patients with diabetes, the direction of the use of cell culture obtained from the pancreas is especially relevant. Given the above, the aim of our study was to study the effect of cell cultures obtained from adipose tissue, bone marrow, and pancreas on the course of experimentally formed insulin-dependent type I diabetes in rats with the aim of developing scientifically based and effective cell therapy methods in veterinary medicine.
Table of Contents
Introduction
Materials and research methods
Research results
CONCLUSION
Objectives and Research Topics
The study investigates the regenerative effects of allogenic stem cell cultures derived from bone marrow, adipose tissue, and the pancreas on experimentally induced type I diabetes in rats to evaluate their potential for clinical cell therapy in veterinary medicine.
- Morphological and phenotypic characterization of stem cell cultures from various sources.
- Assessment of cytogenetic stability of stem cells during in vitro subcultivation.
- Histological evaluation of pancreatic regeneration following stem cell transplantation.
- Quantitative analysis of glucose levels in rats post-transplantation.
- Comparative effectiveness of different stem cell sources in treating alloxan-induced diabetes.
Excerpt from the Book
Red bone marrow stem cell culture
The primary culture of rat red bone marrow stem cells was characterized by morphological heterogeneity. Within a few days after planting, a significant number of attached rounded cells that did not divide could be observed in combination with fibroblast-like cells, which subsequently covered most of the area of culture plastic. Primary bone marrow culture reached 90–100 % confluence on average in 8 days.
In the process of subcultivation, 70–80 % confluence was achieved in 3 days. In passage I, culture heterogeneity was noted. The stem cell culture of rat red bone marrow contained a small number of polygonal cells surrounded by fibroblast-like cells. The morphological heterogeneity of red bone marrow culture stem cells at passages 0 and I can be explained by the fact that the hematopoietic cells contained in bone marrow are able to survive and self-sustain for a long time without a significant increase in their number. Therefore, against the background of a rapid increase in fibroblast-like cells, the percentage of polygonal cells decreases significantly, which we noted at the following passages.
Summary of Chapters
Introduction: Discusses the rising incidence of diabetes in animals, the limitations of current insulin treatments, and the research potential of using stem cell-based regenerative therapies.
Materials and research methods: Details the experimental setup, including animal care, cell culture preparation from bone marrow, adipose tissue, and pancreas, immunophenotyping protocols, and the use of alloxan to model type I diabetes.
Research results: Presents the findings on morphological characteristics, CD marker expression, and cytogenetic stability of the cell cultures, followed by an analysis of the therapeutic impact of transplantation on blood glucose levels and pancreatic tissue structure.
CONCLUSION: Summarizes the study's findings, highlighting that while all cell types showed varying degrees of success, pancreatic cell culture proved most effective in regenerating Langerhans islets and reducing hyperglycemia.
Keywords
Type I diabetes, stem cell culture, bone marrow, adipose tissue, pancreas, Langerhans islets, regenerative medicine, alloxan, cell transplantation, cytogenetic analysis, glucose level, CD markers, veterinary medicine, cell therapy, histology.
Frequently Asked Questions
What is the primary focus of this study?
The research examines the therapeutic effect of transplanting allogenic stem cell cultures derived from bone marrow, adipose tissue, and the pancreas on rats with experimentally induced type I diabetes.
What are the central research areas?
The study covers the morphological and phenotypic characterization of stem cells, their genetic stability in vitro, and their ability to facilitate pancreatic regeneration and glucose regulation in vivo.
What is the main objective of the research?
The goal is to develop scientifically grounded and effective cell therapy methods for treating insulin-dependent diabetes in veterinary practice.
Which scientific methods were employed?
The study utilized cell culture techniques, immunophenotyping to detect CD markers, cytogenetic analysis of metaphase plates, alloxan-induced diabetes modeling, and histological/morphometric analysis of pancreatic tissue.
What is covered in the research results?
This section reports on the growth dynamics, genetic changes (aneuploidy and polyploidy), expression of differentiation markers, and the histological and physiological outcomes of cell transplantation.
Which keywords characterize this work?
Key terms include Type I diabetes, stem cell transplantation, regenerative medicine, Langerhans islets, alloxan, and cytogenetic stability.
Which stem cell source was found to be the most effective?
According to the results, transplantation of the pancreas stem cell culture provided the best therapeutic effect, leading to significant islet neogenesis and a sharper decrease in blood glucose levels.
Did the cell cultures show genetic abnormalities?
Yes, all studied cultures exhibited some level of quantitative chromosomal abnormalities like aneuploidy and polyploidy, but these changes remained within the spontaneous levels characteristic of mammals.
- Citar trabajo
- Vitaly Kovpak (Autor), Anatoly Mazurkevych (Autor), Yuriy Kharkevych (Autor), 2020, The Effect of Allogenic Cell Cultures on Type I Diabetic Rats, Múnich, GRIN Verlag, https://www.grin.com/document/915037
