KC-2617

Ba/F3 STAT3-Luc2-IL21R cell line

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Background

IL21R (Interleukin 21 Receptor) is a Protein Coding gene. Diseases associated with IL21R include Immunodeficiency 56 and Ige Responsiveness, Atopic. Among its related pathways are Cytokine Signaling in Immune system and IL-4 Signaling and its Primary Biological Effects in Different Immune Cell Types. The protein encoded by the IL21R (interleukin 21 receptor) gene is a cytokine receptor for interleukin 21 (IL21), which belongs to the type I cytokine receptor, and has been shown to form a heterodimeric receptor complex with a common γ chain, which is a receptor subunit also shared by the interleukin 2, 4, 7, 9 and 15 receptors. This receptor mediates the growth-promoting signaling of IL21 and is important for the proliferation and differentiation of T cells, B cells, and NK cells. Like other gamma cytokines, IL-21 transduces molecular signals primarily through the Janus kinase and signal transduction and transcriptional activators (JAK-STAT), inositol phosphate 3-kinase (PI3K), and mitosolysis-activated protein kinase (MAPK) pathways. IL-21 induces intense and sustained STAT3 activation, which is critical for its effect on B cell and T cell differentiation. As the converging point of numerous oncogenic signaling pathways, signal transducer and activator of transcription 3 (STAT3) plays a central role in regulating antitumor immune responses. STAT3 is extensively overactivated in both cancerous and non-cancerous cells within the tumor ecosystem and plays an important role in suppressing the expression of key immune activation regulators and promoting the production of immunosuppressive factors. Mutations in human STAT3 are associated with diseases such as immunodeficiency, autoimmunity and cancer. Ba/F3 cell, a murine interleukin-3 dependent pro-B cell line, is a popular system for exploring both kinases and their inhibitors, because some protein kinases can render the Ba/F3 cells to be depended on the activation of the kinases instead of IL-3 supplement, while their inhibitors can antagonize the kinase-dependent growth effects.

Specifications

Catalog Number:
KC-2617
Cell Line Name:
Ba/F3 STAT3-Luc2-IL21R cell line
Host Cell Line:
Mouse Ba/F3 cell line
Description:
Stable Ba/F3-IL21R cell line expressing exogenous luciferase gene under the control of STAT3 signaling pathway
Quantity:
One vial of frozen cells (5×106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% RPMI1640+20% FBS+10% DMSO
Propagation Medium:
RPMI1640+10%FBS+8ng/mL mouse IL-3 +1μg/mL Puromycin+100μg/mL Hygromycin
Selection Marker:
Puromycin, HygromycinB
Morphology:
Mostly single, round (some polymorph) cells in suspension
Subculture:
Split saturated culture 1:10 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 20 hours
Mycoplasma Status:
Negative

Cell Line Generation

Ba/F3-STAT3-Luc2-IL21R cell line was generated using lentivirus methods.

Characterization

Application

Cell Resuscitation

1. Prewarm culture medium (RPMI1640+10%FBS + 8ng/mL mouse IL-3 + 1μg/mL Puromycin + 100μg/mL Hygromycin) 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 1:10 every 3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

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

References

1. Davis MR, Zhu Z, Hansen DM, Bai Q, Fang Y. The role of IL-21 in immunity and cancer. Cancer Lett. 2015 Mar 28.
2. Long D, Chen Y, Wu H, Zhao M, Lu Q. Clinical significance and immunobiology of IL-21 in autoimmunity. J Autoimmun. 2019 May.
3. Zou S, Tong Q, Liu B, Huang W, Tian Y, Fu X. Targeting STAT3 in Cancer Immunotherapy. Mol Cancer. 2020 Sep 24.
4. Hillmer EJ, Zhang H, Li HS, Watowich SS. STAT3 signaling in immunity. Cytokine Growth Factor Rev. 2016 Oct.
5. You L, Wang Z, Li H, Shou J, Jing Z, Xie J, Sui X, Pan H, Han W. The role of STAT3 in autophagy. Autophagy. 2015.
6. Zhao J, Qi YF, Yu YR. STAT3: A key regulator in liver fibrosis. Ann Hepatol. 2021 Mar-Apr.
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