KC-6561

293T-mouse-CCR3-Low Cell Line

64541

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Background of 293T-mouse-CCR3-Low Cell Line

CCR3 (C-C Motif Chemokine Receptor 3) is a Protein Coding gene. The protein encoded by this gene is a receptor for C-C type chemokines. It belongs to family 1 of the G protein-coupled receptors. This receptor binds and responds to a variety of chemokines, including eotaxin (CCL11), eotaxin-3 (CCL26), MCP-3 (CCL7), MCP-4 (CCL13), and RANTES (CCL5). The CCR3 gene is a major research target. The study found that the CCR3 gene is an important target gene for the development of respiratory allergic diseases such as allergic rhinitis and asthma. Diseases associated with CCR3 include Esophagitis and Allergic Rhinitis.

Specifications

Catalog Number	KC-6561
Cell Line Name	293T-mouse-CCR3-Low Cell Line
NCBI/UniProt Accession Number	P51678
Clone Number	8#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous mouse CCR3 gene in low level
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% EMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-mouse-CCR3-Low cell line was generated using a lentiviral vector expressing the mouse CCR3 sequence.

Characterization

Figure 1: Characterization of mouse CCR3 overexpression in the 293T-mouse-CCR3-Low stable clone using FACS.

Figure 2: Characterization of mouse CCR3 in the 293T-mouse-CCR3-Low stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (DMEM+10% FBS +1μg/mL Puromycin) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Tang S, Shu X. [Effect of CCR3 gene on related inflammatory cells in respiratory allergic diseases]. Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2021 Jan 5;35(1):80-84. Chinese. doi: 10.13201/j.issn.2096-7993.2021.01.021. PMID: 33540982; PMCID: PMC10128548.
2. Gangur V, Birmingham NP, Thanesvorakul S, Joseph S. CCR3 and CXCR3 as drug targets for allergy: principles and potential. Curr Drug Targets Inflamm Allergy. 2003 Mar;2(1):53-62. doi: 10.2174/1568010033344453. PMID: 14561176.
3. Rosendahl S, Sulniute R, Persson J, Forsberg S, Häggvik R, Drewsen V, Koskinen Holm C, Kindstedt E, Lundberg P. Lack of CCR3 leads to a skeletal phenotype only in male mice. Biochem Biophys Res Commun. 2022 Sep 10;620:98-104. doi: 10.1016/j.bbrc.2022.06.062. Epub 2022 Jun 24. PMID: 35780587.
4. Amerio P, Frezzolini A, Feliciani C, Verdolini R, Teofoli P, De Pità O, Puddu P. Eotaxins and CCR3 receptor in inflammatory and allergic skin diseases: therapeutical implications. Curr Drug Targets Inflamm Allergy. 2003 Mar;2(1):81-94. doi: 10.2174/1568010033344480. PMID: 14561178.

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.