KC-2007

MC38 human KDR Cell Line

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Background of MC38 human KDR Cell Line

KDR is a tyrosine protein kinase that plays an important role in angiogenesis, vascular development, vascular permeability and embryonic hematopoiesis. KDR also plays an important role in promoting endothelial cell proliferation, survival, migration and differentiation. Diseases associated with KDR include hemangioma, capillary infantile, and vascular disease. Studying the KDR gene may provide a promising new strategy for the treatment of these diseases.

Specifications

Catalog Number	KC-2007
Cell Line Name	MC38 human KDR Cell Line
Host Cell Line	MC38
Description	Stable MC38 cell line expressing exogenous human KDR 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 5μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:8-1:10 every 2-3 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
In Vivo Validation	NA

Cell Line Generation

MC38 human KDR Cell Line was generated using a lentiviral vector expressing the human KDR sequence.

Characterization

Figure 1: Characterization of human KDR overexpression in the MC38 human KDR stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 5μg/ml Puromycin)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 9.0mL 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 1:8-1:10 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Mohammad Rezaei F, Hashemzadeh S, Ravanbakhsh Gavgani R, Hosseinpour Feizi M, Pouladi N, Samadi Kafil H, Rostamizadeh L, Kholghi Oskooei V, Taheri M, Sakhinia E. Dysregulated KDR and FLT1 Gene Expression in Colorectal Cancer Patients. Rep Biochem Mol Biol. 2019 Oct;8(3):244-252. PMID: 32274396; PMCID: PMC7103086.
Whitlock RS, Ebare K, Cheng LS, Fishman DS, Mills JL, Nguyen HN, Nuchtern JG, Ruan W, Smith VE, Patel KA, Fisher KE, Vasudevan SA. Angiosarcoma of the Pancreas in a Pediatric Patient With an Activating KDR-Internal Tandem Duplication: A Case Report and Review of the Literature. J Pediatr Hematol Oncol. 2022 Apr 1;44(3):e751-e755. doi: 10.1097/MPH.0000000000002248. PMID: 34224514.