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Acute lymphoblastic leukemia multidrug resistance cell line

A multi-drug resistance, blast cell technology, applied in the field of cell engineering, can solve the problem of not finding multi-drug resistance cell lines of acute lymphoblastic leukemia

Active Publication Date: 2014-07-23
JIANGSU KEYGEN BIOTECH CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the multidrug-resistant cell line of acute lymphoblastic leukemia used for drug screening

Method used

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  • Acute lymphoblastic leukemia multidrug resistance cell line
  • Acute lymphoblastic leukemia multidrug resistance cell line
  • Acute lymphoblastic leukemia multidrug resistance cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Induction and establishment of acute lymphoblastic leukemia multidrug-resistant cell line MOLT-4 / FU

[0019] The acute lymphoblastic leukemia multidrug-resistant cell line MOLT-4 / FU was established by gradually increasing the concentration of 5-FU and intermittently acting in vitro. The specific steps are as follows:

[0020] (1) Culture medium for human acute lymphoblastic leukemia cell line MOLT-4 (RPMI-1640 culture medium containing 10% calf serum), 5% CO2, 37°C, culture until 70% to 90% adherent When the rate is high, remove the supernatant, add fluorouracil 1000ng / ml impact culture for 1 hour, remove fluorouracil 1000ng / ml culture solution, wash the blank RPMI-1640 culture solution twice, replace the proliferation medium and culture for 72-96h to expand the surviving cells. When the surviving cells expanded to 70%-90% adherence rate, repeat the impact culture with fluorouracil 1000ng / ml culture solution for 1 hour and 9 times, that is, complete 10 times o...

Embodiment 2

[0023] Morphological observation of embodiment 2 drug-resistant strains

[0024] The acute lymphoblastic leukemia multidrug-resistant cell line MOLT-4 / FU in the logarithmic growth phase was taken, and the cell morphology was observed under an inverted microscope. The results were as follows: Image 6 Shown: the cell shape becomes larger and the nucleus shrinks.

Embodiment 3

[0025] The mensuration of embodiment 3 cell growth curve and population doubling time

[0026] Cells were digested, counted, and prepared into a cell suspension with a concentration of 5×103 / mL, and 100 μL of cell suspension was added to each well of a 96-well cell culture plate (500 cells per well); the 6-well cell culture plate was placed at 37°C , cultured in a 5% CO2 incubator for 2d, 4d, 6d, 8d, and 10d respectively; stain the 96-well plate with MTT, add 20 μL of MTT (5 mg / mL) to each well, and continue culturing in the incubator for 4 hours; discard the medium , add 150 μL DMSO to each well to dissolve, shake gently for 10 minutes; λ=490nm, read the OD value of each well with a microplate reader, draw the cell growth curve with time as the abscissa and OD value as the ordinate. For cell growth curves of MOLT-4 and MOLT-4 / FU, see figure 1 .

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Abstract

With a 5-FU concentration gradual increasing and intermittent action in-vitro induction method, the invention establishes an acute lymphoblastic leukemia multidrug resistance cell line MOLT-4 / FU. The cell line is preserved at China General Microbiological Culture Collection Center on December 24th, 2012, and has a preservation number of CGMCC No.7053. The cell line provided by the invention has typical multidrug resistance, and provides an experimental basis for further researches on resistance reversing approaches.

Description

technical field [0001] The invention belongs to the technical field of cell engineering and relates to a human cell line, in particular to an acute lymphoblastic leukemia multidrug-resistant cell line and its establishment method and application. Background technique [0002] Acute lymphoblastic leukemia is a common malignant tumor of the hematopoietic system, and chemotherapy is the main treatment. Although the prognosis of acute leukemia has been greatly improved with the application of new chemotherapeutic drugs, strong multi-drug chemotherapy regimens, and hematopoietic stem cell transplantation, refractory and relapse are still practical problems in the treatment of leukemia. Studies have shown that multidrug resistance is the root cause of chemotherapy failure in most leukemia patients. Multidrug resistance (MDR) refers to resistance to one drug and resistance to other anti-tumor drugs with different structures and different targets. Multi-drug resistance in acute ly...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
Inventor 不公告发明人
Owner JIANGSU KEYGEN BIOTECH CORP LTD
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