• 名称: THP-1(人单核细胞白血病细胞)
  • 货号: CBP60518
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CBP60518

 

 

I. General information

 

Synonyms:

THP1; THP 1; THPI; THP-1(ATCC)

Background:

Derived from the peripheral blood of a 1 year old male with acute monocytic leukaemia at relapse in 1978. The cells can be used for induction of differentiation studies; the cells were described to produce lysozyme and to be phagocytic; carries t(9;11)(p21;q23) leading to MLL-MLLT3 (MLL-AF9) fusion gene. THP-1 cells have Fc and C3b receptors and lack surface and cytoplasmic immunoglobulins. These cells also stain positive for alpha-napthhyl butyrate esterase, produce lysozymes and are phagocytic (both latex beads and sensitised erythrocytes). THP-1 cells can also restore the response of purified T lymphocytes to Concanavlin A, show increased CO2 production on phagocytosis and can be differentiated into macrophage-like cells using for example DMSO.

Species:

Homo sapiens, human

Tissue:

Peripheral blood

Disease:

Acute monocytic leukemia

Gender:

Male, 1-year-old

Morphology:

Round, single cells in suspension, partly in clusters, monocyte

Growth Mode:

Suspension

Doubling Time:

ca. 35-50 hours

DNA Profile:

Amelogenin: X,Y
CSF1PO: 11,13
D13S317: 13,13/8,13
D16S539: 11,12
D5S818: 11,12
D7S820: 10,10
THO1: 8,9.3/8,10
TPOX: 8,11
vWA: 16,16/16,17
Cobioer’s Cell Line Authentication Service

Culture Medium:

RPMI-1650+ 0.05 mM 2-mercaptoethanol+10%FBS
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% FBS+10%DMSO

Photo:

 

Antigen Expression:

HLA A2, A9, B5, DRw1, DRw2.

Receptor Expression:

Complement (C3), expressed

Fc, expressed

Genes Expressed:

Lysozyme,HLA A2, A9, B5, DRw1, DRw2

Immunology:

CD3 -, CD4 +, CD13 +, CD15 +, CD19 -, CD34 -, cyCD68 +, HLA-DR +

Molec. Genetics:

Expression of fusion gene MLL-MLLT3 (MLL-AF9) confirmed by RT-PCR

Viruses:

ELISA: reverse transcriptase negative; PCR: EBV -, HBV -, HCV -, HHV-8 -, HIV -, HTLV-I/II -, MLV -, SMRV -

Comments:

The cells are phagocytic (for both latex beads and sensitized erythrocytes) and lack surface and cytoplasmic immunoglobulin.
Monocytic differentiation can be induced with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA).

For more information, please contact Cobioer (4008-750-250).

 

II. Handling Procedure for Flask Cultures

The flask was seeded with cells grown and completely filled with medium at Cobioer to prevent loss of cells during shipping.
1. Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination. Using an inverted microscope (preferably equipped with phase-contrast optics), carefully check for any evidence of microbial contamination
2. Incubate the flask in an upright position for several hours at 37°C. After the temperature has equilibrated, aseptically remove the entire contents of the flask and centrifuge at 125 x g for 5 to 10 minutes. Remove shipping medium and save for reuse. Resuspend the cell pellet in 10 ml of this medium.
3. From this cell suspension remove a sample for a cell count and viability. Adjust the cell density of the suspension to 4 x 10e5 viable cells/mL in the shipping medium.
4. Incubate the culture, horizontally, at 37°C in a 5% CO2 in air atmosphere. Maintain the cell density of the culture as suggested under the subculture procedure.

 

III. Subculturing Procedure

Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.

Cultures can be maintained by the addition of fresh medium or replacement of medium.
Alternatively, cultures can be established by centrifugation with subsequent resuspension at 2-4x 10e5 viable cells/mL. Subculture when cell concentration reaches 8x10e5 cells/mL.

Do not allow the cell concentration to exceed 1 x 10e6 cells/mL.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: 2 to 3 times per week

 

IV. Cryopreservation Procedure

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
1. Transfer cell suspension to a centrifuge tube and spin at approximately 1000rpmfor 5 to10 minutes.
2. Discard supernatant and resuspend cells in cryopreservation medium at 2x10e6 cells/mL.
3. Transfer the cells into Cryogenic Vials, 1ml/vial.
4. Frozen the cells in cryogenic container (Nalgene #5100-0001).

 

V. Database

Mutation:

Mutation Gene

Zygosity

Gene Sequence

Protein Sequence

CDKN2A

Homozygous

c.1_471del471

p.0?

NRAS

Heterozygous

c.35G>A

p.G12D

TP53

Homozygous

c.520_545del26

p.R174fs*3

ATM

Unknown

c.C>G

p.T935R

For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Expression:

SMAP2, TREM2, SLC2A9
For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Comments:

For more information, please contact Cobioer(4008-750-250).

 

VI. References

1. PubMed=19377474; DOI=10.1038/ng.375; Suzuki H., Forrest A.R.R., van Nimwegen E., Daub C.O., Balwierz P.J., Irvine K.M., Lassmann T., Ravasi T., Hasegawa Y., de Hoon M.J.L., Katayama S., Schroder K., Carninci P., Tomaru Y., Kanamori-Katayama M., Kubosaki A., Akalin A., Ando Y., Arner E., Asada M., Asahara H., Bailey T., Bajic V.B., Bauer D., Beckhouse A.G., Bertin N., Bjorkegren J., Brombacher F., Bulger E., Chalk A.M., Chiba J., Cloonan N., Dawe A., Dostie J., Engstrom P.G., Essack M., Faulkner G.J., Fink J.L., Fredman D., Fujimori K., Furuno M., Gojobori T., Gough J., Grimmond S.M., Gustafsson M., Hashimoto M., Hashimoto T., Hatakeyama M., Heinzel S., Hide W., Hofmann O., Hornquist M., Huminiecki L., Ikeo K., Imamoto N., Inoue S., Inoue Y., Ishihara R., Iwayanagi T., Jacobsen A., Kaur M., Kawaji H., Kerr M.C., Kimura R., Kimura S., Kimura Y., Kitano H., Koga H., Kojima T., Kondo S., Konno T., Krogh A., Kruger A., Kumar A., Lenhard B., Lennartsson A., Lindow M., Lizio M., MacPherson C., Maeda N., Maher C.A., Maqungo M., Mar J., Matigian N.A., Matsuda H., Mattick J.S., Meier S., Miyamoto S., Miyamoto-Sato E., Nakabayashi K., Nakachi Y., Nakano M., Nygaard S., Okayama T., Okazaki Y., Okuda-Yabukami H., Orlando V., Otomo J., Pachkov M., Petrovsky N., Plessy C., Quackenbush J., Radovanovic A., Rehli M., Saito R., Sandelin A., Schmeier S., Schonbach C., Schwartz A.S., Semple C.A., Sera M., Severin J., Shirahige K., Simons C., St Laurent G., Suzuki M., Suzuki T., Sweet M.J., Taft R.J., Takeda S., Takenaka Y., Tan K., Taylor M.S., Teasdale R.D., Tegner J., Teichmann S., Valen E., Wahlestedt C., Waki K., Waterhouse A., Wells C.A., Winther O., Wu L., Yamaguchi K., Yanagawa H., Yasuda J., Zavolan M., Hume D.A., Arakawa T., Fukuda S., Imamura K., Kai C., Kaiho A., Kawashima T., Kawazu C., Kitazume Y., Kojima M., Miura H., Murakami K., Murata M., Ninomiya N., Nishiyori H., Noma S., Ogawa C., Sano T., Simon C., Tagami M., Takahashi Y., Kawai J., Hayashizaki Y.; "The transcriptional network that controls growth arrest and differentiation in a human myeloid leukemia cell line."; Nat. Genet. 41:553-562(2009).

2. PubMed=22460905; DOI=10.1038/nature11003; Barretina J., Caponigro G., Stransky N., Venkatesan K., Margolin A.A., Kim S., Wilson C.J., Lehar J., Kryukov G.V., Sonkin D., Reddy A., Liu M., Murray L., Berger M.F., Monahan J.E., Morais P., Meltzer J., Korejwa A., Jane-Valbuena J., Mapa F.A., Thibault J., Bric-Furlong E., Raman P., Shipway A., Engels I.H., Cheng J., Yu G.K., Yu J., Aspesi P. Jr., de Silva M., Jagtap K., Jones M.D., Wang L., Hatton C., Palescandolo E., Gupta S., Mahan S., Sougnez C., Onofrio R.C., Liefeld T., MacConaill L., Winckler W., Reich M., Li N., Mesirov J.P., Gabriel S.B., Getz G., Ardlie K., Chan V., Myer V.E., Weber B.L., Porter J., Warmuth M., Finan P., Harris J.L., Meyerson M., Golub T.R., Morrissey M.P., Sellers W.R., Schlegel R., Garraway L.A.; "The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity."; Nature 483:603-607(2012).

3. PubMed=23955599; DOI=10.1038/ng.2731; Kon A., Shih L.-Y., Minamino M., Sanada M., Shiraishi Y., Nagata Y., Yoshida K., Okuno Y., Bando M., Nakato R., Ishikawa S., Sato-Otsubo A., Nagae G., Nishimoto A., Haferlach C., Nowak D., Sato Y., Alpermann T., Nagasaki M., Shimamura T., Tanaka H., Chiba K., Yamamoto R., Yamaguchi T., Otsu M., Obara N., Sakata-Yanagimoto M., Nakamaki T., Ishiyama K., Nolte F., Hofmann W.K., Miyawaki S., Chiba S., Mori H., Nakauchi H., Koeffler H.P., Aburatani H., Haferlach T., Shirahige K., Miyano S., Ogawa S.; "Recurrent mutations in multiple components of the cohesin complex in myeloid neoplasms."; Nat. Genet. 45:1232-1237(2013).

4. PubMed=25130606; DOI=10.1016/j.intimp.2014.08.002; Chanput W., Mes J.J., Wichers H.J.; "THP-1 cell line: an in vitro cell model for immune modulation approach."; Int. Immunopharmacol. 23:37-45(2014).
5. PubMed=25485619; DOI=10.1038/nbt.3080; Klijn C., Durinck S., Stawiski E.W., Haverty P.M., Jiang Z., Liu H., Degenhardt J., Mayba O., Gnad F., Liu J., Pau G., Reeder J., Cao Y., Mukhyala K., Selvaraj S.K., Yu M., Zynda G.J., Brauer M.J., Wu T.D., Gentleman R.C., Manning G., Yauch R.L., Bourgon R., Stokoe D., Modrusan Z., Neve R.M., de Sauvage F.J., Settleman J., Seshagiri S., Zhang Z.; "A comprehensive transcriptional portrait of human cancer cell lines."; Nat. Biotechnol. 33:306-312(2015).
6. PubMed=27397505; DOI=10.1016/j.cell.2016.06.017; Iorio F., Knijnenburg T.A., Vis D.J., Bignell G.R., Menden M.P., Schubert M., Aben N., Goncalves E., Barthorpe S., Lightfoot H., Cokelaer T., Greninger P., van Dyk E., Chang H., de Silva H., Heyn H., Deng X., Egan R.K., Liu Q., Mironenko T., Mitropoulos X., Richardson L., Wang J., Zhang T., Moran S., Sayols S., Soleimani M., Tamborero D., Lopez-Bigas N., Ross-Macdonald P., Esteller M., Gray N.S., Haber D.A., Stratton M.R., Benes C.H., Wessels L.F.A., Saez-Rodriguez J., McDermott U., Garnett M.J.; "A landscape of pharmacogenomic interactions in cancer."; Cell 166:740-754(2016).

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