Human Nasopharyngeal Carcinoma Cell Culture Medium 

MCE Human Nasopharyngeal Carcinoma Cell Culture Medium is specially designed to promote the in vitro growth of primary human nasopharyngeal carcinoma cells.

This product utilizes a bicarbonate-based buffering system and includes essential and non-essential amino acids, vitamins, organic and inorganic compounds, hormones, growth factors, and trace elements. Additionally, it contains fetal bovine serum and antibiotics specifically for primary cell culture. This composition provides an optimal in vitro environment for the growth of primary human nasopharyngeal carcinoma cells, enhancing the success rate of primary tumor tissue cell cultures.

-20℃, 1 year

Keep away from light

Pre-experiment Preparation

1. Thaw the culture medium overnight in the 4°C, and then remove it 30 min before use to equilibrate to room temperature.

2. NIH-3T3 cells treated with γ-ray irradiation (40 Gy).

Note: NIH-3T3 cells serve as feeder cells to support the growth of primary human nasopharyngeal carcinoma cells.

Primary Culture of Human Nasopharyngeal Carcinoma Cells

1. Cell Seeding.

1) The initial isolation of small samples of nasopharyngeal carcinoma cell suspension generally does not require counting and can be directly inoculated into 12-well plates.

2) Intraoperative nasopharyngeal carcinoma samples need to pass through a 70 μm cell strainers and be counted. Based on the type of culture dish, refer to the table below to seed primary human neuroblastoma cells. Add γ-irradiated NIH-3T3 feeder cells accordingly. Incubate in a 5% CO₂, 37°C incubator.

2. Medium Change: After seeding the primary cells, avoid moving the culture for 2-3 days to prevent poor cell attachment. If the medium turns yellow but the cells have not yet reached confluence, a half-medium change can be performed.

Note: Primary cells may attach slowly or weakly after initial seeding. To minimize the frequency of observation and handling, it is recommended to check every 2-3 days.

3. Microscopic Observation: During culture, if cells are not fully confluent but feeder cells are insufficient, γ-irradiated NIH-3T3 feeder cells can be added as needed.

Note: The amount of feeder cells added is typically half of the initial seeding amount.

Passage of Primary Human Nasopharyngeal Carcinoma Cells

1. Microscopic Observation: Use a microscope to observe the formation of colonies or the proliferation of neuron-like cells. When the cell confluence reaches 80% - 90%, passaging can be performed.

2. Cell Digestion: Remove the cells from the incubator, discard the old medium, and wash the cells with an appropriate amount of 0.25% trypsin solution for 30 s, then discard the trypsin. Add a fresh aliquot of 0.25% trypsin solution and incubate at 37°C for 5 min.

Note: The dissociation time should not be too long, as excessive dissociation can damage synaptic-like cells and affect cell condition.

3. Microscopic Observation: After dissociation, remove the culture dish and gently tap the side. Observe the cell dissociation status with microscope.

4. Termination of Dissociation: When cells appear rounded and start to detach, add an appropriate amount of culture medium to terminate dissociation. Transfer the cell suspension to a centrifuge tube, centrifuge at 1,500 rpm for 3 min, discard the supernatant and collect the cell pellet.

5. Cell Counting: Resuspend the cell pellet in 1 - 2 mL of culture medium and perform cell count.

6. Cell Seeding: Seed the cell suspension into a culture dish at an appropriate density and add γ-irradiated NIH-3T3 cells. Add culture medium to the required volume and mix thoroughly using the cross-hatch method.

7. Culturing: Place the culture dish in a 5% CO₂, 37°C incubator for culturing.

8. Perform the media change, microscopic observation, and cell feeding operations as described above.

1. This product should be equilibrated to room temperature before use. It is recommended to appropriately aliquot to avoid repeated freeze-thaw cycles.

2. This product is a sterile solution, it is recommended to aseptic operation to avoid contamination.

3. This product contains fetal bovine serum and antibiotics specifically for primary cell culture, so there is no need to add additional serum or dual antibiotics.

4. The experimental samples should be intraoperative or endoscopic samples of nasopharyngeal carcinoma, and the higher the proportion of tumor cells (> 50%), the higher the success rate of primary cell culture.

5. This product is not recommended for culturing small samples (such as circulating tumor cells).

6. To maintain efficacy, it is recommended to avoid prolonged storage of the product at room temperature or in a high-temperature environment.

7. This product is for R&D use only, not for drug, household, or other uses.

8. For your safety and health, please wear a lab coat and disposable gloves to operate.