PD-L1 cell line
PD-L1 Stable Cell Line
Culture conditions:
Cells should be grown at 37 degree C with 5% CO2 using DMEM medium (w/ L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS supplemented with 10% FBS and 1% Pen/Strep, plus 10 ug/ml of Blasticidin.
It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree C water-bath, transfer to a tube containing 10 ml of growth medium without Blasticidin, spin down cells, resuspend cells in pre-warmed growth medium without Blasticidin, transfer resuspended cells to T25 flask and culture in 37 degree C-CO2 incubator.
Leave the T25 flask in the incubator for 1~2 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Blasticidin. Cells should be split before they reach complete confluence.
To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly. To achieve satisfactory results, cells should not be passaged over 16 times.
It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37°C water-bath, transfer to atube containing 10 ml of growth medium without Blasticidin, spin down cells, resuspend cells in pre-warmed growth medium without Blasticidin, transfer resuspended cells to T25 flask and culture in 37°C-CO2 incubator.
Leave the T25 flask in the incubator for 1~2 days without disturbing or changing the medium until cells completelyrecover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Blasticidin. Cells should be split before they reach complete confluence.
To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly. To achieve satisfactory results, cells should not be passaged over 16 times.
Flow Cytometry (FC)
(Detection of human PD-L1 in the CHO-K1/PD-L1 stable cell line by Flow Cytometry [Cell surface staining]. CHO-K1 cells (Green); CHO-K1/PD-L1 cells (Red).)
NCBI and Uniprot Product Information
NCBI Description
This gene encodes an immune inhibitory receptor ligand that is expressed by hematopoietic and non-hematopoietic cells, such as T cells and B cells and various types of tumor cells. The encoded protein is a type I transmembrane protein that has immunoglobulin V-like and C-like domains. Interaction of this ligand with its receptor inhibits T-cell activation and cytokine production. During infection or inflammation of normal tissue, this interaction is important for preventing autoimmunity by maintaining homeostasis of the immune response. In tumor microenvironments, this interaction provides an immune escape for tumor cells through cytotoxic T-cell inactivation. Expression of this gene in tumor cells is considered to be prognostic in many types of human malignancies, including colon cancer and renal cell carcinoma. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Sep 2015]
Uniprot Description
CD274: Involved in the costimulatory signal, essential for T- cell proliferation and production of IL10 and IFNG, in an IL2- dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production. Belongs to the immunoglobulin superfamily. BTN/MOG family. 3 isoforms of the human protein are produced by alternative splicing.
Protein type: Membrane protein, integral
Chromosomal Location of Human Ortholog: 9p24.1
Cellular Component: plasma membrane
Molecular Function: protein binding
Biological Process: cell surface receptor linked signal transduction; immune response; negative regulation of activated T cell proliferation; negative regulation of interferon-gamma production; negative regulation of interleukin-10 production; positive regulation of T cell proliferation; regulation of activated T cell proliferation; response to cytokine stimulus; signal transduction; T cell costimulation
