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Highly aggressive human acute b lymphocytic leukemia cell line with add(11)(q23) chromosomal abnormality

A technology of leukemia cells and B lymphocytes, applied in the fields of biotechnology and microbial animal cell lines, can solve problems such as abnormal chromosome structure, achieve important biological significance and practical value, maintain stable and high proliferation kinetics and genetic characteristics aggressive effect

Inactive Publication Date: 2019-10-29
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most studied is the balanced translocation that occurs in the region where the 11q23 MLL gene is located. The fusion gene of the MLL gene and the partner gene on other chromosome segments often indicates a poor prognosis of the disease, but there are still a considerable number of ALL patients with no MLL gene rearrangement The structural abnormality of the long arm of chromosome 11, including the clonal add(11)(q23) chromosome structural abnormality; research reports, there are 409 known functional genes on the 11q23-q25 segment, including CD3D, THY1, RCK , PLZF and other genes related to the occurrence of blood tumors and tumor suppressor genes such as TSG11, but the role of gene expression or abnormal regulation caused by abnormal rearrangement of non-MLL genes in this region is still unknown in the occurrence and prognosis of ALL. One is that the vast majority of hematological tumor cell lines involving chromosome 11 abnormalities are t(v; 11q23) containing MLL gene rearrangements, while human ALL with structural abnormalities in the long arm of chromosome 11 carrying clonal non-MLL gene rearrangements Cell lines have not been reported yet

Method used

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  • Highly aggressive human acute b lymphocytic leukemia cell line with add(11)(q23) chromosomal abnormality
  • Highly aggressive human acute b lymphocytic leukemia cell line with add(11)(q23) chromosomal abnormality
  • Highly aggressive human acute b lymphocytic leukemia cell line with add(11)(q23) chromosomal abnormality

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Experimental program
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Effect test

Embodiment 1

[0061] Example 1 Isolation and strain establishment of CHH-1 cell strain

[0062] The cells involved in the present invention are all suspended and cultured in the RPMI 1640 cell culture medium containing 10% fetal bovine serum (FBS); the culture temperature is 37° C., and the gas environment is 5% CO 2 / 95% air, humidity is saturated humidity. ; Passage at a ratio of 1:2.

[0063]1) Isolation and inoculation of B-lineage precursor cells from the patient's bone marrow

[0064] ①Put 10ml of the patient's bone marrow fluid obtained by Guchuan into a sterile sodium heparin anticoagulant tube, and separate the patient's bone marrow mononuclear cells with Ficoll human lymphocyte separation medium;

[0065] ②Press 2×10 6 / ml cell density Inoculate the isolated bone marrow mononuclear cells into 24-well cell culture plates, add RPMI 1640 medium containing 50ng / ml recombinant human IL-3 and 50ng / ml recombinant human SCF and 20% FBS, place at 37 °C, the gas environment is 5% CO 2 ...

Embodiment 2

[0075] Example 2 CHH-1 cell line mouse tumorigenicity test and invasion test

[0076] 1) NOD / SCID mouse tumorigenic experiment

[0077] ①The CHH-1 cells in the logarithmic growth phase were divided into 5×10 6 / 200μl / only inoculated subcutaneously in NOD / SCID mice;

[0078] ②Observe the tumorigenicity of the tumor cells at the inoculation site daily, measure the longest diameter and shortest diameter of the tumor mass after tumor formation, and calculate the volume of the tumor mass;

[0079] ③16 days after inoculation, a rice-grain-sized lump could be touched at the subcutaneous cell injection site of the mouse, and a lump protruding from the skin could be observed with the naked eye on the 28th day, and the mouse was sacrificed on the 28th day, and the tumor was separated to make tissue sections for H-E staining and immunohistochemical staining , and after the tumor cells were separated, the karyotype analysis and Ig gene rearrangement detection were performed again;

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Abstract

The invention belongs to the field of microbial animal cell lines, and relates to a new human acute B lymphocytic leukemia cell strain CHH-1. The in vitro isolate of the mononuclear cell of the marrow of a patient with firstly-diagnosed acute B lymphocytic leukemia undergoes cell primary culture to obtain the highly aggressive human acute B lymphocytic leukemia cell strain with add(11)(q23) chromosome abnormality, the preservation number of the cell strain is CGMCC No.11797, and the highly aggressive human acute B lymphocytic leukemia cell strain with add(11)(q23) chromosome abnormality is named as human acute B lymphocytic leukemia cell strain CHH-1, has a same clone source with patient's leukemia cells, can be infinitely and stably passed in vitro, and has the characteristics of clonality add(11)(q23) genetic abnormality, high tumorigenicity and high aggressiveness. The new human acute B lymphocytic leukemia cell strain CHH-1 provides a new good cell model for researches of human acute lymphocytic leukemia with No.11 chromosome long arm structure abnormality and development of biomedicines, and has wide prospect and great practical values in revelation of the pathogenesis of the human acute lymphocytic leukemia and development of medicines.

Description

technical field [0001] The invention belongs to the fields of biotechnology and microbial animal cell lines, and relates to human acute B lymphocytic leukemia, in particular to a highly invasive human acute B lymphocytic leukemia cell line accompanied by add(11)(q23) chromosomal abnormality and its application . Background technique [0002] The prior art discloses that acute lymphoblastic leukemia (ALL) is a group of malignant clonal diseases derived from B-lineage or T-lineage lymphoid precursor cells or lymphoblasts. In 2008, the World Health Organization (WHO) classification system for blood tumors defined it as B- or T-lymphoblastic leukemia / lymphoma (lymphoblastic leukemia / lymphoma), emphasizing the biological characteristics of the disease and downplaying the involved sites . According to statistics, leukemia is one of the top ten high-incidence malignant tumors in my country, with high malignancy and high mortality rate. Especially in children, acute leukemia is th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCC12N5/0694
Inventor 许小平王倩马燕李佩陈波斌庄琳
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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