Human Neuroblastoma Cell Culture Medium 

MCE Human Neuroblastoma Cell Culture Medium is specially designed to promote the in vitro growth of primary human neuroblastoma 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 neuroblastoma cells, enhancing the success rate of primary tumor tissue cell cultures.

-20℃, 1 year

Keep away from light

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. For tumor tissue samples, the higher the proportion of tumor cells (> 50%), the higher the success rate of primary cell culture.

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

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

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

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 neuroblastoma cells.

Primary Culture of Human Neuroblastoma Cells

1. Cell Seeding: 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 the primary cells are seeded, avoid moving the culture for 2-3 days to prevent poor cell attachment. If the medium turns yellow but the cells have not yet confluenced, 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 Neuroblastoma 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 85% - 95%, 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 0.05% trypsin solution for 5 s, then discard the trypsin. Add a fresh aliquot of 0.05% trypsin solution and incubate at 37°C for 2-5 min.

Note: The dissociation time should not be too long, as excessive dissociation can damage synaptic-like cells and affect cell condition. It is recommended to control the dissociation time within 3 min.

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 - 5 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 (seeding ratio as indicated in the table above). 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. For tumor tissue samples, the higher the proportion of tumor cells (> 50%), the higher the success rate of primary cell culture.

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

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

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