DSMZ no.:

Species:

human (Homo sapiens)

Cell type:

essential thrombocythemia

Origin:

established from the peripheral blood of a 71-year-old woman with essential thrombocythemia at megakaryoblastic leukemic transformation in 1995; cells carry the JAK2 V617F mutation and the DNMT3A R882H mutation. Exome and RNA sequence data are available (see Ref 18187 and Exome sequence and RNA-Seq)

Reference(s):

14871, 15030, 18187, 18206

Biosafety level:

1

Permissions and restrictions:

Morphology:

mostly single cells in suspension, some cells grow loosely adherent, 1-3% giant cells; image; image; image

Medium:

80% RPMI 1640 + 20% h.i. FBS

Subculture:

start and cultivate in 24- or 12-well plates for optimal growth; seed out at ca. 0,8 x 10⁶ cells/ml; maintain at 0.5-1.5 x 10⁶ cells/ml; split culture 1:2 every 3-4 days; possibly no cell growth during first week after thawing

Incubation:

at 37 °C with 5% CO₂

Doubling time:

ca. 60-80 hours

Harvest:

maximum density at ca. 1.5 x 10⁶ cells/ml

Storage:

frozen with 70% medium, 20% FBS, 10% DMSO

Mycoplasma:

negative in microbiological culture, PCR assays

Immunology:

CD3 -, CD4 +,CD7 +, CD13 +, CD14 -, CD15 -,CD19 -, CD33 +, CD34 +, CD38 +, CD41 (+), CD42 -, HLA-DR +, GlyA -, CD41+, CD42-; image

Authenticity:

STR analysis according to the global standard ANSI/ATCC ASN-0002.1-2021 (2021) resulted in an authentic STR profile of the reference STR database.

Cytogenetics:

human flat-moded near-diploid karyotype with 8% polyploidy; 46-47<2n>XX, +13, -14, +21, +mar, add(4)(p15), der(5)t(5;19)(q11;q13), der(7)t(7;14)(q31;q11), del(9)(p11p24)ins(9;?21)(p11;p11p12), add(9)(q11)t(9;22)(p24;q12), der(13)del(13)(q11-q13)t(13;21)(q34;q11-12), der(13)t(13;21)(q34;q11-12), ins(16;9)(q22;?p23p24), der(19)t(9;19)(p24;q13), ider(?21;9)(p10p13;p24), der(21)r(21;9)(p1?1q1?1;p24p2?3); carries multiple rearrangements effecting 9p24 amplification; resembles published karyotype

Viruses:

PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, SMRV -

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