KC-2128

MC38-FOLH1-Middle-Cell-Line

23446

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Background of MC38-FOLH1-Middle-Cell-Line

FOLH1 belongs to the type II transmembrane glycoprotein of the M28 peptidase family. This protein acts as glutamate carboxypeptidase on various alternative substrates, including folate and neuropeptide N-acetyl-1-aspartate-1-glutamate, and is expressed in many tissues such as the prostate, central and peripheral nervous systems, and kidneys. The mutation of this gene may be related to impaired intestinal absorption of dietary folate, leading to low blood folate levels and subsequently causing hyperhomocysteinemia. The expression of this protein in the brain may be associated with many pathological conditions related to glutamate excitotoxicity. In the prostate, this protein is upregulated in cancer cells and is used as an effective diagnostic and prognostic indicator for prostate cancer.

Specifications

Catalog Number	KC-2128
Cell Line Name	MC38-FOLH1-Middle-Cell-Line
Host Cell Line	MC38
Description	Stable MC38 clone expressing exogenous FOLH1 gene in middle level
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 1:6-1:8 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	Yes

Cell Line Generation

MC38-FOLH1-Middle-cell-line was generated using a lentiviral vector expressing the FOLH1 sequence.

Characterization

Figure 1: Characterization of FOLH1 overexpression in the MC38-FOLH1-Middle stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 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 1:6-1:8 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

Plichta KA, Graves SA, Buatti JM. Prostate-Specific Membrane Antigen (PSMA) Theranostics for Treatment of Oligometastatic Prostate Cancer. Int J Mol Sci. 2021 Nov 9;22(22):12095. doi: 10.3390/ijms222212095. PMID: 34829977; PMCID: PMC8621856.
Hong X, Mao L, Xu L, Hu Q, Jia R. Prostate-specific membrane antigen modulates the progression of prostate cancer by regulating the synthesis of arginine and proline and the expression of androgen receptors and Fos proto-oncogenes. Bioengineered. 2022 Jan;13(1):995-1012. doi: 10.1080/21655979.2021.2016086. PMID: 34974814; PMCID: PMC8805960.