Human acute B lymphocyte leukemia cell line and application thereof

A technology of acute lymphocytes and B lymphocytes, applied in the detection of programmed cell death, animal cells, tumors/cancer cells, etc., can solve the problems of relapsed and refractory leukemia, poor prognosis, and refractory patients

Active Publication Date: 2018-07-06
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although, in the past 50 years, the prognosis of children with acute lymphoblastic leukemia has made great progress, the 5-year survival rate has increased from 10% in the 1960s to 90%, and the 5-year event-free survival rate has exceeded 80%, but there are

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  • Human acute B lymphocyte leukemia cell line and application thereof
  • Human acute B lymphocyte leukemia cell line and application thereof
  • Human acute B lymphocyte leukemia cell line and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Isolation and strain establishment of human acute lymphoblastic leukemia cell line HXEX-ALL1

[0035] 1) Isolation and inoculation of naive B lymphocytes from the patient's bone marrow

[0036] Extract 3ml of the patient's bone marrow, anticoagulate with heparin, and separate the patient's bone marrow mononuclear cells with Ficoll-Hypaque lymphocyte separation medium (TBD Tianjin Haoyang Biological Products Company); press 6×10 6 Cells / ml density were resuspended in RPMI 1640 medium (Gibco Company) containing 10ng / ml IL-3 (Sigma Company), 20% fetal bovine serum (Thermo Company), 37°C, saturated humidity, 5% CO 2 Incubator cultivation.

[0037] After inoculation of bone marrow mononuclear cells, the medium was changed 48 hours to remove cell debris. Cells were counted every other day. After 7 days, the number of cells began to increase steadily, and the number of dead cells decreased. Primary cultured cells were established.

[0038] According to the cell de...

Embodiment 2

[0048] Example 2: Identification of Growth and Biological Characteristics of Human Acute B Lymphoblastic Leukemia Cell Line HXEX-ALL1

[0049] 1) Observation of cell morphology

[0050] Take the HXEX-ALL1 cells in the logarithmic growth phase and observe the living cell morphology under an inverted microscope, as shown in figure 1 As shown, the cells are growing in suspension, round, and the shape is relatively uniform. HXEX-ALL1 cells in the logarithmic growth phase were taken, fixed with routine smears, stained with Swiss-Jimusa, and the cell morphology was observed under an upright microscope, as shown in figure 2 As shown, the cells are round, with a diameter of 11-15 µm, the ratio of nucleoplasm to plasma is large, mitosis can be seen, the nucleus is round, and nucleoli can be seen.

[0051] HXEX-ALL1 cells in the logarithmic growth phase were taken, routinely fixed and embedded, and the ultrastructure of the cells was observed with a transmission electron microscope, ...

Embodiment 3

[0058] Establishment of humanized acute lymphoblastic leukemia animal model with cell line HXEX-ALL1

[0059] Tumor formation experiment in NOD / SCID mice: HXEX-ALL1 cells in the logarithmic growth phase were divided into 5×10 6 Each / 100µl / mouse, inoculated subcutaneously in the bare area of ​​female Balb / c (nu / nu) mice (6 weeks old, weighing 14-16g); observe the tumor formation at the inoculation site every other day, and the results are as follows: Figure 6 As shown, 5 days after inoculation, the size of rice grains could be touched under the skin of nude mice. The tumor grew rapidly after 11 days of inoculation, and the size of the tumor was measured every 3 days. It can be seen that the volume of the tumor increased from 248 mm on the 11th day. 3 1707mm grown to day 20 3 14 days later, the tumor block was taken to make a cell suspension, and immunophenotyping and STR detection confirmed that the cells isolated from the tumor block were from the same source as the HXEX-ALL...

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Abstract

The invention provides a human acute B lymphocyte leukemia cell line HXEX-ALL1. The human acute B lymphocyte leukemia cell line is preserved in China Center for Type Culture Collection (CCTCC) on October 25, 2017, and the preservation number is CCTCC NO: C2017232. The cell line is a primary line-established human acute B lymphocyte leukemia cell line and is a human acute B lymphocyte leukemia cellline with high tumorigenicity. The cell line provides a novel cell model for researches of relapse and refractory human acute lymphocyte leukemia and researches and development of biomedicines; the cell line can be provided for establishing a humanized acute lymphocyte leukemia animal model, an acute lymphocyte leukemia generation mechanism and relapse and refractory mechanism research platform,an acute lymphocyte leukemia novel drug research and development and screening platform, an acute lymphocyte leukemia immunological and microbiological research platform and the like. The human acuteB lymphocyte leukemia cell line HXEX-ALL1 provided by the invention has a wide prospect and relatively great application value for exploring a human acute lymphocyte leukemia pathogenesis, a relapse and refractory mechanism and drug research, development and application.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to a human acute B lymphocytic leukemia cell line and application thereof. Background technique [0002] Acute lymphoblastic leukemia (ALL) is a group of malignant clonal diseases derived from B-line or T-line lymphoid precursor cells or lymphoblastoid cells. It is the most common malignant tumor in childhood and seriously threatens children's health. life. In recent years, the incidence of acute lymphoblastic leukemia in children has been on the rise. B-acute lymphoblastic leukemia (B-ALL) accounts for approximately 85% of childhood ALL. Although, in the past 50 years, the prognosis of children with acute lymphoblastic leukemia has made great progress, the 5-year survival rate has increased from 10% in the 1960s to 90%, and the 5-year event-free survival rate has exceeded 80%, but there are still 10-20 % of patients are refractory, relapsed, and the prognos...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02A01K67/027
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/03C12N5/0694C12N2503/02G01N33/5011G01N2500/10G01N33/5052
Inventor 顾玲朱易萍张鸽杨荣张研乐孙舒雯
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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