Human Acute Myeloid Leukemia Cell Culture Medium 

MCE Human Acute Myeloid Leukemia Cell Culture Medium is specially designed to promote the in vitro growth of primary human acute myeloid leukemia 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. The higher the proportion of human acute myeloid leukemia cells (> 30%), 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.

Sample Separation

1. Collect Blood Cells: Mix the peripheral blood or bone marrow sample from the anticoagulant tube and transfer it into a 15 mL centrifuge tube. Centrifuge at 1,500 rpm for 5 min. Discard the plasma and retain the cell pellet.

2. Dilute Blood Cells: Add PBS (1×) to the blood cell pellet to dilute it. The final volume of the cell suspension should be 9 mL.

3. Peripheral Blood Cell Separation: Prepare a 15 mL centrifuge tube and add 6 mL of human peripheral blood cell separation fluid. Slowly add this to the diluted cell suspension. Centrifuge at 400 g, with an acceleration rate of 2 and a deceleration rate of 0, at 25°C for 30 min.

4. Collect Leukocytes: After centrifugation, the centrifuge tube will separate into four layers from top to bottom: PBS layer, the creamy white leukocyte layer, separation fluid layer, and red blood cell layer. Prepare another 15 mL centrifuge tube with 5 mL of PBS (1×) and use a pipette to collect the leukocyte layer into the tube containing PBS.

5. Wash Leukocytes: Gently mix the leukocyte suspension, centrifuge at 1,500 rpm for 3 min at room temperature. Discard the supernatant and collect the cell pellet.

6. Lysis: Add 6 mL of red blood cell lysis buffer (MCE catalog number: HY-K3010) to resuspend the cell pellet. Incubate at 4°C for 15-20 min. Centrifuge at 1,500 rpm for 3 min at room temperature, discard the supernatant, and collect the cell pellet.

Note: Gently invert the tube once during the lysis process to mix.

7. Recover Leukocytes: Add 1-5 mL of complete culture medium to resuspend the cell pellet.

8. Cell Counting and Seeding: Perform a live cell count on the cell suspension. Based on the cell count, seed the cells into culture plates as per the table below. Place the plates in a 5% CO₂, 37°C incubator for culture.

Cell Passage

1. Microscopic Examination: Observe cell morphology under a microscope. Passage the cells when the suspension reaches 80% - 90% confluency.

2. Collect Cells: Transfer the cell suspension into a 15 mL centrifuge tube. Centrifuge at 1,500 rpm for 3 min at room temperature. Discard the supernatant and collect the cell pellet.

3. Counting and Seeding: Add 1 - 2 mL of complete culture medium to resuspend the cell pellet. Perform a live cell count and seed the cells into culture plates as per the table above. Place the plates in a 5% CO₂, 37°C incubator for culture.

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 higher the proportion of human acute myeloid leukemia cells (> 30%), 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.