KC-1047

A549-OS8 Cell Line

21439

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Background of A549-OS8 Cell Line

The OS8 antigen is a cell surface protein identified as a potential tumor-associated antigen. It is highly expressed in certain cancers, most notably osteosarcoma (as suggested by its name), making it a candidate for targeted immunotherapy research. Its expression profile in malignancies compared to limited expression in normal tissues positions it as a promising target for the development of antibody-based therapies, such as monoclonal antibodies or CAR-T cells, aimed at treating solid tumors.

Specifications

Catalog Number	KC-1047
Cell Line Name	A549-OS8 Cell Line
Clone Number	NA
Host Cell Line	A549
Description	Stable A549 cell line expressing exogenous OS8 gene
Quantity	Two vials 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% McCoy’s 5a + 20% FBS + 10% DMSO
Propagation Medium	McCoy's 5a+10%FBS+500µg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 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 22 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

A549-OS8 Cell Line was generated using a lentiviral vector expressing the OS8 sequence.

Characterization

Figure 1: Characterization of OS8 overexpression in the A549-OS8 stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (McCoy's 5a+10%FBS+500µg/mL Hygromycin B)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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% McCoy’s 5a + 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

Latouche, Jean-Baptiste; Sadelain, Michel (2000). "Induction of human cytotoxic T lymphocytes by artificial antigen-presenting cells". Nature Biotechnology. 18 (4): 405–409.
Perica, Karlo; Kosmides, Alyssa K; Schneck, Jonathan P (2015). "Linking form to function: Biophysical aspects of artificial antigen presenting cell design". Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 1853 (4): 781–790.