Cell lines were obtained from the German Collection of Microorganisms and Cell Cultures (Brunswick, Germany). All cells but retinal pigment epithelial (RPE) were cultured in RPMI-1640 medium (Invitrogen, Grand Island, NY) supplemented with penicillin/streptomycin and 10% fetal bovine serum. RPE cells were cultured in DMEM (Invitrogen, Grand Island, NY) medium supplemented with penicillin/streptomycin and 10% fetal bovine serum. Suspension and adherent cells were maintained at a density of 0.5–1 million cells/ml or 50–90% confluence, respectively, at 37 ºC in a humidified atmosphere with 5% CO2. Jurkat cells engineered to express GFP were generated by retroviral transduction as described.23 (link)–24 (link) Cell counts were measured using the Neubauer hemocytometer (Hausser Scientific, Horsham, PA), according to manufacturer’s instructions.
Protocol detail
Cell Culture Protocols for Adherent and Suspension Cells
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Atmosphere
Cell cultures
Cell lines
Cells
Epithelial cells
Fetal bovine serum
Jurkat cells
Penicillin
Retinal pigment
Retroviral
Streptomycin
Corresponding Organization :
Other organizations : Kettering University, Boston Children's Hospital, Harvard University, Memorial Sloan Kettering Cancer Center, Cornell University
Top 5 similar protocols
Variable analysis
independent variables
- Cell line type (e.g., Jurkat, RPE)
- Cell count
- Cell culture medium (RPMI-1640 for all cells except RPE, DMEM for RPE)
- Supplementation of culture medium (penicillin/streptomycin and 10% fetal bovine serum)
- Cell culture conditions (37 °C, 5% CO2, humidified atmosphere)
- Cell density (0.5-1 million cells/ml for suspension cells, 50-90% confluence for adherent cells)
- Jurkat cells engineered to express GFP were used as a positive control to demonstrate the retroviral transduction protocol
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!