Atypical chronic myeloid leukemia cell line and preparation method thereof

A technology for chronic myeloid cells and leukemia cells, which is applied in the field of human atypical chronic myeloid leukemia cell line NT-1 and its preparation, and can solve the problems of little treatment experience and no satisfactory treatment method.

Active Publication Date: 2014-03-26
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, there is little experience in the treatment of aCML and no satisfactory treatments

Method used

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  • Atypical chronic myeloid leukemia cell line and preparation method thereof
  • Atypical chronic myeloid leukemia cell line and preparation method thereof
  • Atypical chronic myeloid leukemia cell line and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Obtainment of human atypical chronic myelogenous leukemia cell line NT-1

[0047] 1. Primary culture

[0048] Extract 2ml of the patient's bone marrow and anticoagulate with sodium citrate; dilute the whole blood with an equal volume of PBS; take an equal volume of Ficoll-Hypaque lymphocyte separation medium (purchased from Shanghai Pufei Company) and add it to a 15ml conical centrifuge tube. Rise to room temperature (18-25°C); tilt the centrifuge tube at an angle of 45 degrees, draw the diluted blood sample with a glass pipette, and slowly spread it on the lymphocyte separation medium along the tube wall at 1 cm above the layered liquid surface, pay attention Do not disturb the interface of the liquid layer; place it in a horizontal centrifuge after balancing, and centrifuge at 2000r / min at room temperature for 20 minutes; take out the centrifuge tube smoothly, and the junction of the plasma layer and the layered liquid is a turbid gray-white layer, rich in ...

Embodiment 2

[0059] Example 2: Identification of growth and genetic characteristics of human atypical chronic myelogenous leukemia cell line NT-1

[0060] 1. Observation of cell morphology

[0061] Transfer a certain number of cells into a 1.5ml centrifuge tube (EP tube), centrifuge at 1500r / min for 5 minutes, discard 2 / 3 of the supernatant, blow the remaining cell suspension evenly, smear, and perform Wright's staining, the cells are of different sizes (20-30 microns), round or oval; nuclei are round or oval, part of the nucleus is folded, twisted, and sunken, part of the nuclear chromatin is fine-grained, part of it is loosely reticular, irregular nucleoli can be seen, part of Cytoplasm without granules, some cytoplasm with fine sand-like granules, cytoplasm is grayish blue, and vacuoles are occasionally seen (see figure 1 ).

[0062] 2. Growth curve of NT-1 cell line

[0063] Make a standard curve:

[0064] (1) First count the number of cells in the prepared cell suspension with a c...

Embodiment 3

[0080] Example 3: Animal experiment of human atypical chronic myelogenous leukemia cell line NT-1

[0081] Six 4-week-old Balb / c nude mice (3 males and 3 females, purchased from the Animal Experiment Center of Nantong University) were collected to collect NT-1 cells in the logarithmic growth phase, and the cell concentration was adjusted to 2×10 8 / ml, each nude mouse was subcutaneously injected with 100 microliters on the right back, and the tumor growth at the injection site was observed. After feeding for 1 month, tumors of various sizes formed at the injection site, about 0.7×0.8~1.1×1.8cm 2 , the nude mice were sacrificed by cervical dislocation, the solid tumor tissues of the nude mice were separated, routinely fixed, embedded in paraffin, sectioned, and stained with HE (see Figure 5 A). The solid tumor tissue of nude mice was isolated under aseptic conditions, cut into pieces with ophthalmic scissors, and filtered through 100-mesh and 200-mesh filters after grinding. ...

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Abstract

The invention belongs to the technical field of leukemia cell lines, and provides an atypical chronic myeloid leukemia cell line NT-1 of which the collection number is CCTCC NO:C2013103, and the atypical chronic myeloid leukemia cell line NT-1 is collected in the China center for type culture collection on August 1, 2013. The invention further provides a preparation method and application of the leukemia cell line NT-1. A new ideal is provided for diagnosis and treatment of atypical chronic myeloid leukemia, and the atypical chronic myeloid leukemia cell line and the preparation method have significance on early diagnosis and drug screening of the atypical chronic myeloid leukemia.

Description

technical field [0001] The invention belongs to the technical field of leukemia cell lines, and in particular relates to a human atypical chronic myelogenous leukemia cell line NT-1 and a preparation method thereof. Background technique [0002] Leukemia cell lines play an important role in the basic research of leukemia. The use of cell lines for in vitro experiments and the construction of animal models can not only deeply study the mechanism of tumor occurrence and development at the molecular and gene levels, but also have important implications for early clinical diagnosis, drug therapy, etc. Screening and treatment are of great importance. [0003] Leukemic cell lines reported in literature at home and abroad and commonly used in leukemia diagnosis and treatment research include: human promyeloid acute leukemia cell line HL-60, human chronic myeloid leukemia cell line K562, etc. [0004] Human atypical chronic myelogenous leukemia (full name in English, aCML for short...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02A61K35/12A61P35/02A61K35/13
Inventor 刘红钱娟刘杰
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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