Human monocytic leukemia cell line THP-1

Human monocytic leukemia cell line THP-1

【THP1; O-THP-1; Tohoku Hospital Pediatrics-1】

Cell identification

STR identification has been passed

Cell source

National Resource Library

Cell background

Established from peripheral blood of a 1-year-old boy with relapsed acute monocytic leukemia (AML) in 1978. This cell can engulf milk droplets and activated red blood cells, without surface and cytoplasmic immunoglobulin. Mononuclear cell differentiation can be induced by using phorbol TPA.

Note: The NRAS mutation was incorrectly indicated as p.Gly12Ser in PubMed=9379676.

Cell characteristics

1) Source: Acute monocytic leukemia, monocytes

2) Form: Monocyte, suspended

3) Content:>1x106 cell count

4) Specification: Packaging in T25 bottles or 1mL cryovials

5) Purpose: For scientific research purposes only

Cultivation conditions

1) Prepare RPMI-1640 (recommended awcell-0002) culture medium; High quality fetal bovine serum, 10%; 0.05 mM β - qiu based ethanol (cell culture grade recommendation: awcell-8211); Double antibody, 1%.

2) Reference passage ratio: During passage, control the cell density at 2-4 × 10 ^ 5 cells/mL, and proceed with passage when the cells grow to 8-10 × 10 ^ 5 cells/mL.

3) Reference fluid change frequency: Change the fluid during passage.

4) Cultivation conditions: Gas phase: air, 95%; Carbon dioxide, 5%. Temperature: 37 degrees Celsius, humidity in the incubator is 70% -80%.

5) Cryopreservation solution: 95% culture medium, DMSO 5% (After using this cryopreservation solution, according to our experience, the survival rate after recovery is about 50%. After recovery, an additional 7-10 days of cultivation are required to restore growth)

L Precautions

[Precautions]:

This cell is a suspension cell, and based on our cultivation experience and customer feedback, using the 'half exchange method' during passaging is more advantageous for the cell state. Therefore, our library recommends that you use the 'half exchange method' for passaging.

1. After receiving the cells shipped in 15ml centrifuge tubes provided by us, please do not collect the cells by centrifugation. Instead, prepare two new T25 culture bottles, mix the cells evenly, and transfer them into two new T25 culture bottles. Add 10ml of culture medium and place them in a 37 ℃ incubator.

2. If you have received cells shipped in T25 culture bottles, please collect the cells by centrifugation first, then resuspend the cells in 12ml of * culture medium prepared according to the instructions and blow evenly before transferring them into two new T25 culture bottles for cultivation.

3. THP-1 suspension cells. The cell prefers gentle treatment during cultivation and minimizes centrifugation to prevent damage to the cell. When changing the medium, do not replace the entire culture medium. It is recommended to use the 'semi liquid change method' for passage, and add cell culture medium to dilute the cells to maintain density.

4. Cells are sensitive to serum quality, and we suggest that you use imported high-quality serum for cultivation. Do not inactivate FBS. If the cell growth is slow, try using other FBS or temporarily increase the FBS concentration to 20%

5. β - qiu based ethanol (cell culture grade) needs to be added to the culture medium of the cell. If not added, it may affect the cell state.

The stability of β - qiu based ethanol is limited, and it cannot be added to the culture medium for long-term storage. β - mercaptoethanol should be added during each passage or liquid addition.

β - qiu based ethanol (2-qiu based ethanol) is added to lymphocyte or other cell cultures because the amino acid cysteine in FBS used in the culture medium is in short supply, while cysteine is abundant. Some cells, such as T cells, are unable to transport cysteine into the cytoplasm and must convert it into cysteine. β - qiu based ethanol reduces cysteine to a form that can be transported by cells, and then converts it into cysteine required for cell growth. β - qiu based ethanol is also a reducing agent that can break down many toxic metabolites produced by cultured cells, thereby improving the environment around the cells.

6. This cell is sensitive to cell density, so please pay attention to maintaining the cell density within an appropriate range during cultivation and passage (please refer to the cell manual for details).

7. The recovery rate of the cell after cryopreservation is relatively low. Please increase the cell count as appropriate during cryopreservation

Normally, cell growth can be maintained by adding * culture medium to the growing cells. Every 7 days, the cells can be centrifuged and resuspended in a newly prepared culture medium to complete the complete cell replacement.

ATCC's explanation on maintaining cell viability of thp-1 cells

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Frequent cell counting is the best method for monitoring THP-1. These cells should be cultured every 2 to 3 days by simply adding a small amount of fresh culture medium to the flask (to give them conditioned medium). You don't need to centrifuge the cells and subculture them every time. When cultured in our laboratory, cells can usually expand into larger flasks with more culture medium by the second day. When the cell density reaches 8 × 10 ^ 5 live cells/ml, passage is required. Cell density exceeding 1 × 10 (6) live cells/ml is not allowed

1. Discard the culture supernatant and rinse the cells 1-2 times with PBS that does not contain calcium or magnesium ions.

2. Add 0.25% (w/v) trypsin 0.53 mM EDTA to culture bottles (1-2mL for T25 bottles and 2-3mL for T75 bottles), digest in a 37 ℃ incubator for 1-2 minutes (digestion time can be appropriately extended for difficult to digest cells), and then observe the digestion of cells under a microscope. If most of the cells become round and fall off, quickly take them back to the operating table, tap the culture bottle a few times, and add 3-4ml of culture medium containing 10 μ S to terminate digestion.

3. Gently mix and aspirate, centrifuge at 1000RPM for 3-5 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly. Divide the cell suspension into new T25 bottles in a ratio of 1:2, add 6-8ml of new * culture medium prepared according to the instructions to maintain the growth vitality of the cells, and subsequently subculture according to the actual situation in a ratio of 1:2~1:5.

3) Cell cryopreservation: It is recommended to freeze a batch of cell seeds during the first 3 generations of culture after receiving the cells for subsequent experimental use.

4) The transport medium (infusion medium) cannot be used to culture cells anymore. Please use the newly prepared * medium according to the instructions for cell culture conditions to culture cells. After receiving the cells, it is recommended to subculture them for the first time in a T25 culture bottle at a ratio of 1:2.

Transportation form

Low temperature: (1) Transport in 1mL cryovials packaged with dry ice. After receiving, store in a -80 degree freezer overnight and transfer to liquid nitrogen or directly resuscitate. If you find that the dry ice has evaporated completely, the cryovial cap has fallen off, is damaged, or the cells are contaminated, please contact us immediately.

At room temperature: (2) T25 bottles of revived surviving cells will be shipped at room temperature and processed according to the procedures for cell reception upon receipt.

Biosafety

1. All animal cells are considered to have potential biological hazards and must be operated in a secondary biosafety platform. Please pay attention to protection, and all waste liquids and containers that have come into contact with these cells must be sterilized before disposal.

2. It is recommended to always use protective gloves, clothing, and a face mask when reviving frozen cells. Attention: The cryotube immersed in liquid nitrogen will leak and gradually fill with liquid nitrogen. When thawing, the conversion of liquid nitrogen into gas phase may cause the container to explode or the lid to be blown off with dangerous force, resulting in flying debris and causing personal injury.