Hematopoietic progenitor kinase (HPK1) is a member of a family of mitogen-activated protein kinase kinase kinase kinase (MAP4K) of Ste20 serine/threonine kinases, which is predominantly expressed in hematopoietic cells and has been shown to be a negative immune regulator of T-cell receptor (TCR) and B-cell signaling.
Cell Line Name:
Jurkat HPK1 HIBIT 2B2 Cell Line
Host Cell Line:
Human Jurkat cell line
One vial of frozen cells (5X106 per vial)
Stable in culture over a minimum of 10 passages
Drug screening and biological assays
70% RPMI1640+20% FBS+10% DMSO
Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL
37 °C with 5% CO2
Liquid nitrogen immediately upon receiving
Approximately 26 hours
Cell Line Generation
Jurkat-HPK1-HIBIT cell line was generated using CRISPR-Cas9 methods.
Characterization using PCR sequencing and WB
1. Prewarm culture medium (RPMI1640 + 10%FBS) 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 1:4-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL.
1. Prepare the freezing medium (70% RPMI-1640 + 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
1. Huizhen Ge, Lizeng Peng, Zhou Sun, Huanxiang Liu , Yulin Shen and Xiaojun Yao. Discovery of Novel HPK1 Inhibitors Through Structure-Based Virtual Screening. ORIGINAL RESEARCH published: 14 March 2022 doi: 10.3389/fphar.2022.850855
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