Induced pluripotent stem cells (iPSC)
Service Brief
Induced pluripotent stem cells are the four transcription factor Oct4, Sox2, Klf4, c-Myc into terminally differentiated cells, which will become the reprogramming of somatic cells into embryonic stem cell like state of pluripotent cells, called induced pluripotent stem cells. Induced pluripotent stem cells have very similar biological characteristics of cells and embryonic stem, due to the unique biological properties of embryonic stem cells, provides an efficient research platform for the study of cell differentiation regulation, disease models, drug screening and clinical treatment.
Service content:
1. induced pluripotent stem cell lines (retroviruses, mRNA, episomal)
2. induced pluripotent stem cell markers immunostaining (Nanog, Oct4, Sox2, SSEA3, SSEA4, TRA1-60, TRA1-81)
3. differentiation of multipotent stem cells in vitro (EB differentiation, three germ layer marker staining)
4. differentiation ability (immunodeficient mice, teratoma formation, immunohistochemistry)
5. chromosome karyotype identification, G band
Primary cell culture
Service brief:
The animal tissue is removed from the body, by various enzymes (usually trypsin), chelating agent (EDTA) or mechanical processing method, dispersed into single cells and cultured in appropriate medium, cell survival, growth and reproduction, a process known in primary culture.
Service content:
Primary cell isolation, culture and identification (identification can be provided by flow cytometry, immunocytochemistry, RT-PCR, protein immunoblotting and other methods).
Continous cell culture
Service brief:
Subculture is one of the conventional conservation methods for cell culture, and is also the basis of all cell biology experiments. When cells in culture flasks covered after the need to dilute the minutes into the bottle, the cells can continue to grow, this process is called. A large number of cells needed for experiments can be obtained by subculture. The passage of cells should be carried out under strict aseptic conditions, and each step should be carefully and carefully sterilized.
Service content:
The identification and management of cell lines (or strains) are provided according to customer requirements.
Stable cell line screening
Service brief:
The exogenous plasmid DNA is integrated into the host cell chromosome, which enables the host cell to express the target protein for a long time, which is called stable transfection. To establish a stable cell line is to screen target cells, select the corresponding drugs according to the resistance markers contained in different gene vectors and screen them selectively with G418.
Service content:
1. cell culture, transient transfection experiments and drug concentration screening;
2. transient transfection screening;
3. construction and identification of polyclonal cell lines (RT-PCR, WB);
4. screening and identification of monoclonal cell lines (R, PCR, WB).
Cell transfection
Service brief:
The selection of transfection technology has a great influence on the transfection results. Many transfection methods need to optimize the proportion of DNA and transfection reagents, the number of cells, the culture and detection time, etc.. The main transfection methods were liposome transfection, electroporation and viral infection.
Service content:
1. liposome transfection
Cationic liposome surface with positive charge, and phosphate of nucleic acids by electrostatic interaction, DNA molecular package in the formation of DNA lipid complexes, can be adsorbed on the surface of the cell membrane is negatively charged, and then through the fusion or endocytosis into cells. Liposome transfection is suitable for transfection of DNA into suspension or adherent cells. It is one of the most convenient transfection methods in the laboratory, and its transfection rate is higher than that of calcium phosphate method. Since liposomes are cytotoxic to cells, the transfection time is generally not more than 24 hours. Commonly used cell types: COS-7, BHK, NIH3T3, Hela and so on.
2. electroporation transfection
The electric current can reversibly breakdown the cell membrane, form an instantaneous water pathway or a small pore on the membrane, and cause the DNA molecule to enter the cell. This method is electroporation. When the transfection efficiency of some liposomes is very low or cannot be transferred, electroporation is suggested to be transfected by electroporation. In general, high field strength kills 50%-70% cells. Our company has developed an electric transfer agent for cell death, which greatly reduces cell death rate and improves transfection efficiency.
3. virus infection
The cell lines transfected with liposome and electroporation could not be successfully transfected. It is recommended that the virus be infected with this method. This method can be used for rapid detection of 100% infection, and the detection rate is high.
