Introduction into mammalian cell culturing methods
Hungary(HuMSIRC)-University of Pecs,Pecs
Department of Medical Biology and Central Electron Microscopic Laboratory
Hajnalka Ábrahám MD, PHD
Katalin Kiss MD
English, German
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No Yes Yes No No No No No No No No
Type of Research Project
- Basic science
What is the background of the project?
Cell culturing techniques are nowdays routinely performed, essential methods of any types of molecular biological laboratories in both basic and applied research fields. Working with either cell lines or cells primaly originated from animal or human tissues, the maintenance and manipulation of cells under these arteficial conditions in dishes require basicly general laboratory activities. Our laboratory has been providing both instrumental and human resources needed to complete these techniques for more than 20 years. Several graduate and undergraduate students, even researchers have become users during this time.
What is the aim of the project?
In this project the student can meet several basic mammalian cell culturing methods and achieve enough routine to work later on his/her own.
What techniques and methods are used?
- preparation of different serum complemented culturing media - preparation of general culturing solutions - plating of cell lines - preparation of primary mammalian cultures maintenance of primary cultures and cell lines short and long term storage techniques (freezing methods) - cell counting - elimination or prevention of common infections (mainly Mycoplasma contamination) - use of inverted phase contrast microscope and cell culture laboratory centrifuges - transfection techniques - preparation of cell lysates for nucleic acid (genomic DNA or RNA) analysis - preparation of cell lysates for protein analysis - preparation of samples for microscopic staining or immunocytochemical methods
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 will be done under supervision
What are the tasks expected to be accomplished by the student?
- plating of cell lines: thrawing up frozen cells and then plating them onto a special culturing dish, flask; chemical pretreatment of dish surface if required; - maintenance of primary cultures and cell lines: keeping cells in incubation chambers, daily observation of cells under inverted phase contrast microscope, change of culturing medium in required timepoints; - cell counting for further biochemical analysis: collection of plated cells via trypsinisation into tubes, centrifugation, counting cells in haemocytometer under microscope, replating cells of required number into fresh culturing cells - short and long term storage techniques (freezing methods): collection of plated cells via trypsinisation into tubes, centrifugation, - resuspending cells in special freezing solutions and cool down of cells at storage temperature (-80°C or liquid nitrogen)
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
optional participation in actually ongoing courses provided by our department
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
knowledge of basic cell biology usage of ordinary bench light microscope
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- Graduated students (less than 6 months)
- Pre-Medical students from the American-British system
- Students in biomedical fields
- Dental medicine students (IADS members)
- Best practices in cell culture: an overview Baust JM; Buehring GC; Campbell L; Elmore E; Harbell JW; Nims RW; Price P; Reid YA; Simione F. In Vitro Cell Dev Biol Anim. 2017 Sep; 53(8):669-672. doi: 10.1007/s11626-017-0177-7. Epub 2017 Aug 14. Review. PMID:28808859