KC-3065

K562-HLA-A1101-Cell-Line

26887

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Background of K562-HLA-A1101-Cell-Line

HLA-A belongs to the HLA class I heavy chain paralogues. HLA-A is a heterodimer consisting of a heavy chain and a light chai. The heavy chain is anchored in the membrane. HLA-A plays a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen so that they can be recognized by cytotoxic T cells. They are expressed in nearly all cells. HLA-A is a Protein Coding gene. Diseases associated with HLA-A include Severe Cutaneous Adverse Reaction and Birdshot Chorioretinopathy. Among its related pathways are SARS-CoV-2 Infection and Infectious disease.

Specifications

Catalog Number	KC-3065
Cell Line Name	K562-HLA-A1101-Cell-Line
Host Cell Line	K562
Description	Stable K562 cell line expressing exogenous HLA-A1101 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 + 300µg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

K562-HLA-A1101 Cell Line was generated using a lentiviral vector expressing the HLA-A1101 sequence.

Characterization

Figure1: Characterization of HLA-A overexpression in K562 HLA A1101 stable clone using FACS.

Figure2: Characterization of HLA-A expression in K562 cell using FACS.

Figure3: Characterization of K562 HLA-A1101 Cell Line stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI164 supplemented with 10% FBS and 300µg/mL Hygromycin B)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI164 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1. Arce-Gomez B, Jones EA, Barnstable CJ, Solomon E, Bodmer WF (February 1978). The genetic control of HLA-A and B antigens in somatic cell hybrids: requirement for beta2 microglobulin. Tissue Antigens. 11 (2): 96–112. doi:10.1111/j.1399-0039.1978.tb01233.x. PMID 77067.
2. de Campos-Lima PO, Levitsky V, Brooks J, et al. (April 1994). T cell responses and virus evolution: loss of HLA A11-restricted CTL epitopes in Epstein-Barr virus isolates from highly A11-positive populations by selective mutation of anchor residues. J. Exp. Med. 179 (4): 1297–305. doi:10.1084/jem.179.4.1297. PMC 2191457. PMID 7511684.

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.