KC-3023

K562-B2M-HLA-A0201 Cell Line

26865

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Background of K562-B2M-HLA-A0201 Cell Line

B2M encodes beta-2-microglobulin, a serum protein that associates with the major histocompatibility complex class I heavy chain on the surface of nearly all nucleated cells. It has a predominantly beta-pleated sheet structure and belongs to the beta-2-microglobulin family. The protein is found in the extracellular region, on the cell surface, in the plasma membrane, and in extracellular exosomes.HLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play 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.

Specifications

Catalog Number	KC-3023
Cell Line Name	K562-B2M-HLA-A0201 Cell Line
NCBI/UniProt Accession Number	NM_004048, NM_002116.7
Clone Number	3#
Host Cell Line	K562
Description	K562 cell line stably expressing exogenous B2M-HLA-A0201 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS +300 μg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Suspension
Subculture	Split saturated culture 1:3-1:6 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 40 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

K562-B2M-HLA-A0201 cell line was generated using a lentiviral vector expressing the B2M-HLA-A0201 sequence.

Characterization

Figure 1: Characterization of B2M-HLA-A0201 overexpression in K562-B2M-HLA-A0201 stable clones using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 300 μg/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 9.0 mL 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 1:3-1:6 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Bataille S, Fernandez C, Zink JV, Brunet P, Berland Y, Burtey S. The Case | A hip fracture in a hemodialysis patient. Pathologic right-hip fracture from β2-microglobulin amyloidosis. Kidney Int. 2013 Jun;83(6):1211-2. doi: 10.1038/ki.2012.436. PMID: 23728017.
de Melo AC, Lucena E, de Oliveira DCM, Viola JPB. Frequency of HLA-A*02:01 in the Brazilian population and its impact on uveal melanoma systemic treatment. Oncologist. 2024 Aug 5;29(8):e1098-e1099. doi: 10.1093/oncolo/oyae112. PMID: 38785402; PMCID: PMC11299924.

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.