Myelodysplastic syndrome-transformed leukocyte line stably expressing green fluorescent protein

Abstract

The invention belongs to the field of microbial animal cell lines. Designing a myelodysplastic syndrome-transformed leukocyte strain stably expressing green fluorescent protein and its establishment method and application. The present invention uses the human myelodysplastic syndrome-transformed leukocyte line SKM-1 as the maternal cell, and performs transfection by a lentivirus carrying the GFP gene. GFP-positive single-cell clones were obtained by limiting dilution method, and the expanded cultured cells were screened in mice by subcutaneous injection. After tumorigenesis, the tumor mass was separated, and after continuous culture, stable expression of green fluorescent protein was obtained. Myelodysplastic syndrome-transformed leukocyte line SKM‑1 / GFP. The cell line has no difference in morphology and growth characteristics from the parental cells, the growth curve has no difference from the parental cells, can stably express GFP, and has tumorigenicity. It can be further used to establish mouse animal models and provide a platform for MDS minimal residual disease and other preclinical research.

Description

technical field [0001] The invention belongs to the fields of biotechnology and microbial animal cell lines, and relates to a myelodysplastic syndrome-transformed leukocyte strain, in particular to a myelodysplastic syndrome-transformed leukocyte strain stably expressing green fluorescent protein and its preparation method and application. Background technique [0002] The prior art discloses that myelodysplastic syndrome (myelodysplastic syndrome, MDS) is a group of diseases of abnormal proliferation of clonal cells, characterized by ineffective hematopoiesis, pathological hematopoiesis and high risk of transformation to acute myeloid leukemia (AML), wherein, About 20%-30% of MDS patients progress to AML. According to reports, in the United States, MDS is the most common tumor of myeloid origin, and the incidence rate of men is higher than that of women, with an average of every 10 5 In every American, 3-4 people suffer from MDS; with the increase of age, the incidence of ...

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Problems solved by technology

Compared with primary AML (de novo AML), patients with MDS-transformed AML (sAML) are older and difficult to treat, and often cannot be treated with allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and intensive chemotherapy, and their prognosis is poor. Good; studies also show that with the development of demethylation drugs such as decitabine and azacitidine, breakthroughs have been made in the treatment of MDS, but these drugs have limited therapeutic effects on high-risk MDS and sAML

Due to the lack of effective treatment methods in clinical practice, it is difficult to remove leukemia cells in the body, which ultimately threatens the lives of patients; however, there is currently a lack of effective and reliable in vivo and in vitro research platforms at home and abroad, which seriously restricts the basic and preclinical research of MDS

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