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In vitro Maintenance of Germ Stem Cell Microenvironment in Testis
Turkey (TurkMSIC) - Hacettepe University, Ankara
Hacettepe University, Faculty of Medicine, Department of Histology and Embryology, Sihhiye 06100 Ankara/Turkey
Prof. Dr. Petek Korkusuz
Tutor(s) Prof. Dr. Petek Korkusuz
Type of Research Project
- Basic science
What is the background of the project?
In childhood cancers, 46% infertility rate is seen in male patients because of the damage in germ cells during the chemotherapy and radiotherapy. It is not possible to collect mature sperms from those patients; since the spermatogenesis in testes does not start until the puberty and there is no sperm production. The long-term culture of spermatogonial stem cells is challenging by using classical methods. Organ culture setup are recent systems, which can provide suitable conditions for modeling the stem cell niches in the body. The Turkish Society of Histology and Embryology, founded in 1990, organizes annual meetings in order to further knowledge of its respective fields, and to strengthen the connections between other branches of science. It also aims to prevent malpractice of its respective fields.
What is the aim of the project?
The project aims primarily to increase the proliferation and subsequent differentiation of spermatogonial stem cell after enlargement of their pool in in vitro culture conditions by using special co-culture platform.
What techniques and methods are used?
What techniques and methods are used? Mention the steps/stages in this project. (Provide examples and avoid abbreviations; which tools will be used in general in this project?) Flow cytometry: The sample is loaded onto the cytometer. The cells in the sample are surrounded by a physiological buffer called sheath fluid. While flowing down, the cells are exposed to a laser beam at the interrogation point. Based on the structures located within the cell, the light is scattered. The lateral and vertical diversion of the beam can be associated within each structure within the cell. In addition, the laser will excite the fluorophores in the cell, creating fluorescent emissions. Immunohistochemistry: Antigens are marked with antibodies, which are then marked with peroxidase or alkaline phosphatase. The resulting complex emits fluorescent lights. Histochemistry: Antigens are reacted with specific chemicals. The resulting product will emit fluorescent lights. Stem cell isolation. ELISA (Enzyme Linked Immunosorbent Assay): Immobilization of the antigen of interest, can be accomplished by direct adsorption to the assay plate or indirectly via a capture antibody that has been attached to the plate. The antigen is then detected either directly (labeled primary antibody) or indirectly (labeled secondary antibody). The most powerful ELISA assay format is the sandwich assay. This type of capture assay is called a “sandwich” assay because the analyte to be measured is bound between two primary antibodies – the capture antibody and the detection antibody. The sandwich format is used because it is sensitive and robust.
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
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
What are the tasks expected to be accomplished by the student?
The following tasks are expected to be accomplished by the student: -Cell culture preparation. -Subculturing of the cells. -Mesenchymal stem cell isolation from bone marrow. -Tissue culture preparation. -Learning about immunohistochemistry and the common antigens presented in spermatogonial stem cell. -Learning about the respective antibodies of common antigens mentioned above. -Learning about the strucutre and functions of antibodies.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will be introduced to the topic of the project by giving related books and articles by research colleagues in the lab and supervisor. Also, he/she will be able to attend seminars and conferences about the concerned subject.
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 presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No specific skills are required.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Pre-Medical students from the American-British system
- Kose; S.; N. Yersal; S. Onen ve P. Korkusuz (2018). "Comparison of Hematopoietic and Spermatogonial Stem Cell Niches from the Regenerative Medicine Aspect." Adv Exp Med Biol. doi: 10.1007/5584_2018_217.
- Komeya; M.; H. Kimura; H. Nakamura; T. Yokonishi; T. Sato; K. Kojima; K. Hayashi; K. Katagiri; H. Yamanaka; H. Sanjo; M. Yao; S. Kamimura; K. Inoue; N. Ogonuki; A. Ogura; T. Fujii ve T. Ogawa (2016). "Long-term ex vivo maintenance of testis tissues producing fertile sperm in a microfluidic device." Sci Rep 6: 21472 doi: 10.1038/srep21472
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