CHO-K1/CD80 cell line穩(wěn)轉(zhuǎn)細胞株

I. INTRODUCTION
Catalog Number: M00614
Cell Line Name: GS-C1/CD80
Gene Synonyms: B7; B7-1; B7.1; BB1; CD28LG; CD28LG1; LAB7
Expressed Gene: Codon Optimized from NM_005191.3; no expressed tags
Host Cell: GS-C1
Quantity: Two vials of frozen cells (1×106 per vial)
Stability: 20 passages
Application: in vitro functional assay
Freeze Medium: 95% complete growth medium, 5% DMSO
Complete Growth Medium: F12K, 10% FBS
Culture Medium: F12K, 10% FBS, 4 μg/ml Puromycin
Mycoplasma160: Negative
Functional Performance: For Ipilimumab, Signal / Background (S/B) > 3
Storage: Liquid nitrogen immediately upon receipt
II. BACKGROUND
Cluster of Differentiation 80 (also CD80 and B7-1) is a protein found on activated B cells and monocytes that
provides a costimulatory signal necessary for T cell activation and survival. It is the ligand for two different proteins
on the T cell surface: CD28 (for autoregulation and intercellular association) and CTLA4 (for attenuation of
regulation and cellular disassociation). CD80 works in tandem with CD86 to prime T cells.
IV. THAWING AND SUBCULTURING
Thawing Protocol
1. Remove the vial from liquid nitrogen tank and thaw cells quickly in a 37°C water-bath.
2. Just before the cells are completely thawed, decontaminate the outside of the vial with 70% ethanol and
transfer the cells to a 15 ml centrifuge tube containing 9 ml of complete growth medium.
3. Pellet cells by centrifugation at 200 x g force for 5 min, and remove the medium.
4. Resuspend the cells in complete growth medium.
5. Transfer the cell suspension to a 10 cm dish with 10 ml of complete growth medium.
6. Grow the cells in incubator with 37°C, 5 %CO2.
7. Add antibiotic in the following day.
Sub-culturing Protocol
1. Remove the culture medium from cells.
2. Wash cells with PBS (pH=7.4) to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 ml of 0.05% (w/v) Trypsin- EDTA (GIBCO, Cat No. 25300) solution into 10 cm dish and observe the
cells under an inverted microscope until cell layer is dispersed (usually within 3 to 5 minutes).
Note: To avoid cells clumping, do not agitate the cells by hitting or shaking the dish while waiting for the cells
detach. If cells are difficult to detach, please place the dish in 37°C incubator for ~2 min.
4. Add 6.0 to 8.0 ml of complete growth medium into dish and aspirate cells by gently pipetting.
5. Centrifuge the cells at 200 x g force for 5min, and remove the medium.
6. Resuspend the cells in culture medium and add the cells suspension to new culture dish.
7. Grow the cells in incubator with 37°C, 5 %CO2.
Subcultivation Ratio: 1:3 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days

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