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The effect of mesenchymal stem cell niche on sperm for experimental diabetic rat models
Turkey (TurkMSIC) - Celal Bayar University, Manisa
Histology and embriyology Department Uncubozköy, MCBU Saglık Bilimleri kampusu, Mimar Sinan Blv. No: 173, 45030 Yunusemre/Manisa
Prof. Dr. Ibrahim TUGLU
Prof. Dr. Ibrahim TUGLU
Type of Research Project
- Basic science
What is the background of the project?
Please, provide the background of the project (Provide information about the project; min. 10 lines, max. 25 lines) Unexplained male infertility accounts for 30-40% of men with abnormal sperm parameters.The causes of spermatogenic defects in infertile patients are multifactorial. The therapeutic effect of stem cells in reproductive health is being studied. As an important source of fat-induced mesenchymal stem cells (FIMSC) has been increasingly used effectively. Improvement problems and sperm anomalies in diabetes-related infertility process is an important problem in medicine and studies are ongoing. A standard method of mimicking Type 2 diabetic status is to grow cells with high glucose or high insulin in the medium.When the sperm cells are thawed, they must return with the least possible damage and be capable of maintaining the survival of the species. However, cell structure and function damage resulting from a sudden drop in temperature in this freeze-thaw process is related to changes in membrane permeability and cell skeleton structure. Sperm motility and spermoocyte fusion capacity may decrease after thawing. Sperm taken from healthy animals with TESE (Testicular Sperm Extraction) will be subjected to diabetic damage in high glucose environment and left to freeze and thaw. After thawing, we aim to minimize the damage caused by freezing thawing in experimental diabetic model by applying KH (Potassium Humate) and niche in culture medium.
What is the aim of the project?
To develop a new method for creating more close-to-live in-vitro models of diabetic sperm cells, thereby increasing the ease with which urological and endocrinological research in this regard can be conducted in the future.
What techniques and methods are used?
Mesenchymal stem cells (MSCs) are well known adult stem cells present in many tissues such as adipocytes, chondrocytes, osteoblasts, and neurons. Many isolations and characterization methods have emerged to apply MSCs in the clinical applications. We use mesenchymal stem cell isolation, culture and expansion in this project. Immunocytochemistry is a common laboratory technique that is used to anatomically visualize the localization of a specific protein or antigen in cells by use of a specific primary antibody that binds to it. TUNEL [Terminal deoxynucleotidyl transferase dUTP(Deoxyuridine Triphosphate) nick end labeling] is a method for detecting apoptotic DNA fragmentation, widely used to identify and quantify apoptotic cells, or to detect excessive DNA breakage in individual cells .The assay relies on the use of terminal deoxynucleotidyl transferase (TdT), an enzyme that catalyzes attachment of deoxynucleotides, tagged with a fluorochrome or another marker, to 3'-hydroxyl termini of DNA double strand breaks. It may also label cells having DNA damaged by other means than in the course of apoptosis.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The following tasks will be expected to be accomplished by the student: -Learning Cell culture protocols passaging, thawning and freezing. - Usage Light Microscope and finding cell area with the help of tutor - Learning Immunohistochemistry techniques - Usage and applying of the Cell Staining types with the help of tutor - Preaparing scientific reports and presentation -Learning mesenchymal stem cell isolation techniques - Learning TUNEL Methods
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
There are many different subjects open for understanding methods.The subject will be taught once per week by tutor Prof. Dr. Ibrahim TUGLU -Cell Cycles -Mesenchymal stem cell isolation protocols -Isolated organ -Cell Culture Protocols -Immunochemistry techniques - TUNEL assay and protocols
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No spesific skills are required.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- • Hsiao; C. H.; Ji; A. T. Q.; Chang; C. C.; Chien; M. H.; Lee; L. M.; & Ho; J. H. C. (2019). Mesenchymal stem cells restore the sperm motility from testicular torsion-detorsion injury by regulation of glucose metabolism in sperm. Stem cell research & therapy; 10(1); 270. doi.org/10.1186/s13287-019-1351-5
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