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The GPC1 gene (Glypican-1) is located in the 2q37.3 region of the human genome and encodes a cell-surface heparan sulfate proteoglycan comprising 558 amino acids. Through its heparan sulfate chains, the protein binds to the extracellular matrix and growth factors (such as Hedgehog and Wnt), thereby regulating processes including cell signaling, proliferation, and differentiation. GPC1 is highly expressed in various tumors—including pancreatic cancer, breast cancer, and non-small cell lung cancer—and contributes to tumor progression by promoting angiogenesis and epithelial-mesenchymal transition (EMT); notably, its expression levels in the blood demonstrate high sensitivity and specificity for the early diagnosis of pancreatic cancer, making it a promising biomarker. Furthermore, GPC1 is associated with neurological disorders (such as spinal muscular atrophy) and plays a role in prion protein conversion. Research currently focuses on its potential as a therapeutic target in oncology—encompassing the development of targeted nanomedicines and the investigation of signaling pathway regulatory mechanisms—thereby offering new avenues for precision cancer therapy.

Figure 1. Recombinant GPC1 CHO stably expressing GPC1.

Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed  CHO-K1 Human GPC1 Cell complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.

Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.

2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.

Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.