KC-2456

M-07e-PD1-Cell-Line

24098

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Background of M-07e-PD1-Cell-Line

PD-1, programed cell death 1 protein, is a receptor belonging to immunoglobulin superfamily which is expressed primarily on activated T cells. NK cells, B cells, and monocytes. PD-1 interaction with its ligands, PD-L1 and PD-L2, leads to downregulation of T cell responses, including T cell proliferation and cytokine production, and limits immune-destruction of tissues. The interaction between PD-1 and its ligands, thus plays a role in maintaining the balance between immune activation and tolerance, potentially including tumor tolerance.

Specifications

Catalog Number	KC-2456
Cell Line Name	M-07e-PD1-Cell-Line
Host Cell Line	Human M-07e cell line
Description	Stable M-07e cell line expressing exogenous human PD1 gene
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+10ng/mLGM-CSF
Selection Marker	Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture 1:3-1:5 every 3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 38 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

M-07e human PD1 Cell Line was generated using a lentiviral vector expressing the human PD1 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640+10%FBS+10ng/mLGM-CSF) in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:3-1:5 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Sunshine, J. & Taube, J. M. PD-1/PD-L1 inhibitors. Current Opinion in Pharmacology 23, 32ÿ38 (2015).
Boussiotis, V. A. Molecular and Biochemical Aspects of the PD-1 Checkpoint Pathway. N Engl J Med 375, 1767ÿ 1778 (2016).
Topalian, S. L. et al. Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N Engl J Med 366, 2443ÿ2454 (2012).

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.