Human KIT Stable Cell Line-Ba/F3

Name: Human KIT Stable Cell Line-Ba/F3
SKU: CSC-RO1109M
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-RO1109M

Host Cell : Ba/F3 Size : >1x10⁶ frozen cells/vial

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Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Cat. No.	CSC-RO1109M
Description	Ba/F3-KIT cell line is engineered to stably overexpress human KIT.
Target Gene	KIT
Gene Species	Homo sapiens (Human)
Alias	C-Kit, CD117, MASTC, PBT, SCFR
Host Cell	Ba/F3
Host Cell Species	Mus musculus (Mouse)
Applications	Drug screening and biological assays
Biosafety Level	2
Size	>1x10^6 frozen cells/vial, 1 mL
Stability	Stable in culture over a minimum of 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Growth Conditions	37 °C, 5% CO₂
Storage	Liquid nitrogen
Shipping	Dry ice

Thawing & Subculturing Instructions	1. Thaw cells by gently swirling in a 37°C water bath. To limit contamination, do not submerge the O-ring and cap. 2. When cells are ~70% thawed (~1 min), transfer the vial into a biosafety cabinet, and wipe the surface with 70% ethanol. Allow tube to dry completely. 3. Transfer the cells gently into a 15 mL conical tube containing 10 mL of pre-warmed culture medium (without antibiotic selection marker). Centrifuge cells at ~125 x g for 5~7 min. 4. Remove supernatant without disturbing the pellet, and resuspend cells in 1 mL culture medium (without antibiotic selection marker). Transfer cells to a 6-well plate containing ~2 mL pre-warmed growth medium (without antibiotic selection marker) or a T25 flask containing 5 mL pre-warmed culture medium (without antibiotic selection marker). 5. Incubate the culture at 37°C with 5% CO2. 6. Subculture: split saturated culture 1:4 ~ 1:6 every 3 days; seed out at about 1~3 x 10^5 cells/mL.
Freeze Medium	Frozen with 70% medium, 20% FBS, 10% DMSO
Growth Properties	Suspension, round
Freezing Instructions	Cells are recommended to generate additional frozen stocks at early passages. Frozen stocks should be preserved in a designated cryopreservation medium or in 70% RPMI 1640 + 20% FBS + 10% DMSO (without antibiotic selection marker). 1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO, without antibiotic selection marker) 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 x g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3 x10^6 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.

Mycoplasma	Negative
Format	One frozen vial containing millions of cells
Storage	Liquid nitrogen
Safety Considerations	The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship	Dry ice

Gene Name	KIT v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog [ Homo sapiens ]
Gene Symbol	KIT
Synonyms	PBT; SCFR; C-Kit; CD117
Gene Description	v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog
GeneID	3815
Uni ProtID	P10721
mRNA Refseq	NM_000222.2
Protein Refseq	NP_000213.1
Chromosome Location	4q11-q12
Function	ATP binding; cytokine binding; metal ion binding; protease binding; protein binding; protein homodimerization activity; protein tyrosine kinase activity; receptor signaling protein tyrosine kinase activity; stem cell factor receptor activity; transmembrane receptor protein tyrosine kinase activity;
Pathway	Acute myeloid leukemia, organism-specific biosystem; Acute myeloid leukemia, conserved biosystem; Adaptive Immune System, organism-specific biosystem; C-MYB transcription factor network, organism-specific biosystem; Constitutive PI3K/AKT Signaling in Cancer, organism-specific biosystem; Cytokine-cytokine receptor interaction, organism-specific biosystem; Cytokine-cytokine receptor interaction, conserved biosystem;
MIM	164920

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Background

Case Study

Applications

Q & A

Customer Reviews

The KIT proto-oncogene, also known as c-Kit or CD117, encodes a receptor tyrosine kinase (RTK) critical for various cellular processes, including proliferation, survival, and differentiation of hematopoietic progenitor cells and melanocytes. Aberrant activation of this gene is closely associated with the occurrence and progression of multiple tumors.

The Ba/F3 cell line, initially derived from murine lymphoid cells, has been extensively utilized in the functional characterization of oncogenes and signaling pathways due to its dependency on interleukin-3 (IL-3) for growth. Its characteristics render it an ideal cellular model for investigating novel oncogenes.

The establishment of the human KIT stable cell line-Ba/F3 represents a significant advancement. By stably transfecting human KIT cDNA into Ba/F3 cells, the study of KIT-mediated signaling and its implications in cellular physiology and pathology became feasible. This cell line's establishment has provided valuable insights into the molecular mechanisms underlying KIT-associated diseases, such as gastrointestinal stromal tumors (GISTs) and acute myeloid leukemia (AML).

Through these studies, we can better comprehend the role of KIT in normal cellular functions and disease development, thereby laying a theoretical foundation for the diagnosis and treatment of related diseases.

The maintenance of adult long-term hematopoiesis relies on the preservation of hematopoietic stem/progenitor cells (HSPC) within specialized niches in the bone marrow (BM). These niches tightly regulate the equilibrium between quiescence, self-renewal, and differentiation of HSPC. Researchers utilized the Human KIT Stable Cell Line-Ba/F3 to investigate the intrinsic responsiveness of HSPC to niche factors. By expressing membrane-targeted C3G (C3G-F), a Rap1 GTP/GDP exchanger, they observed enhanced proliferation and differentiation of HSPC in response to stem cell factor (SCF) and thrombopoietin (TPO). This prolonged activation of c-Kit receptor and downstream signaling through SCF ligation sheds light on the crucial role of basal Rap1 activation in maintaining HSPC in bone marrow niches, crucial for long-term adult hematopoiesis.

Figure 1. Increased basal Rap1GTP level in Human KIT Stable Cell Line-Ba/F3 leads to prolonged stem cell factor (SCF)‐mediated c‐Kit receptor activation and downstream signaling. Protein expression and cell surface c‐Kit were examined, revealing altered levels. Effects of SCF stimulation on cell viability and migration were assessed. Signaling dynamics were evaluated through immunoprecipitation and immunoblotting. (Imai T, et al., 2019)

1. Oncology Research: Utilize Human KIT Stable Cell Line-Ba/F3 for investigating targeted therapies against gastrointestinal stromal tumors (GISTs), examining drug efficacy, and understanding mechanisms of drug resistance.

2. Drug Screening: Employ Human KIT Stable Cell Line-Ba/F3 to screen novel compounds for their potential as KIT inhibitors, facilitating the discovery of new anti-cancer drugs.

3. Signaling Pathway Studies: Investigate intracellular signaling pathways activated by KIT receptor using Human KIT Stable Cell Line-Ba/F3, shedding light on cellular processes involved in cancer development and progression.

4. Resistance Mechanism Analysis: Study acquired resistance mechanisms to KIT-targeted therapies in GISTs by employing Human KIT Stable Cell Line-Ba/F3, aiding in the development of strategies to overcome drug resistance.

5. Preclinical Models: Utilize Human KIT Stable Cell Line-Ba/F3 as a preclinical model for evaluating the efficacy and safety of potential therapeutic agents targeting KIT, facilitating the translation of research findings into clinical applications.

Customer Q&As

Why were Ba/F3 cells chosen for establishing the stable cell line expressing human KIT?

A: Ba/F3 cells were chosen for establishing the stable cell line expressing human KIT due to their lack of endogenous KIT expression and their responsiveness to KIT activation. Ba/F3 cells offer a suitable background for studying KIT signaling and drug sensitivity.

What measures were taken to verify and maintain the stability and expression level of human KIT in the Ba/F3 stable cell line?

A: Verification and maintenance of stability and expression level of human KIT in the Ba/F3 stable cell line involved stable transfection, clonal selection, and validation of expression using techniques such as qPCR and western blot analysis. Continuous culture under appropriate conditions ensured stable expression.

Please explain the functional characterization of human KIT in the Ba/F3 stable cell line, emphasizing its kinase activity and response to KIT inhibitors.

A: Functional characterization of human KIT in the Ba/F3 stable cell line focused on its kinase activity and response to KIT inhibitors. This included assessing the phosphorylation of downstream signaling molecules such as STAT5 and ERK, as well as evaluating sensitivity to KIT inhibitors like imatinib using cell viability assays.

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Customer Reviews

Valuable resource for understanding oncogenesis

An invaluable resource! The Human KIT Stable Cell Line has significantly enhanced my research capabilities, offering valuable insights into KIT-mediated signaling networks and potential therapeutic strategies for KIT-associated diseases.

French

2023-12-26

Reliable tool for KIT-targeted therapies

Impressive reliability. This cell line surpasses expectations, serving as a robust platform for studying KIT-targeted therapies and potential interventions for KIT-driven malignancies. Its stable expression simplifies experimental workflows, facilitating efficient data collection and analysis, and accelerating discoveries in hematology and cancer research.

Germany

2023-07-27

Facilitating hematopoiesis studies

With stable KIT expression, I can investigate signaling pathways downstream of KIT and KIT-mediated cellular processes with confidence, advancing our understanding of hematopoietic disorders and oncogenesis. The Human KIT Stable Cell Line in Ba/F3 cells ensures consistent expression of KIT, providing reliable results in my research on hematopoiesis and cancer.

United States

2020-05-22

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Human KIT Stable Cell Line-Ba/F3

Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Background

Case Study

Applications

Q & A

Customer Reviews

Why were Ba/F3 cells chosen for establishing the stable cell line expressing human KIT?

What measures were taken to verify and maintain the stability and expression level of human KIT in the Ba/F3 stable cell line?

Please explain the functional characterization of human KIT in the Ba/F3 stable cell line, emphasizing its kinase activity and response to KIT inhibitors.

Valuable resource for understanding oncogenesis

Reliable tool for KIT-targeted therapies

Facilitating hematopoiesis studies

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