BTHS inducible multi-functional stem cells, preparation method thereof, differential culture medium and purpose

Abstract

The invention discloses BTHS inducible multi-functional stem cells, a preparation method thereof, a differential culture medium and a purpose. The preparation method comprises the steps of taking 150-200 milliliters of urine of a BTHS patient, and performing centrifugation at 2000 revolution per minute for 10 minutes to obtain urine cells; after the cells are counted, with the quantity of 3.5-4*10<5>, performing inoculation to a 6-hole culture board in which matrigel is incubated at 37 DEG C for an hour or above; after passage, culturing third-generation urine cells to reach 80% of convergence, infecting sendai viruses containing four transcription factors of OCT4, SOX2, KLF4 and c-MYC, and performing reprogramming induction; after 15 days of viral infection, starting to generate clone regiment like cells, through manually selecting clone regiment cells, performing transferring to a new 24-hole culture board in which the matrigel is incubated, and performing culture; and after repeatedpassage, completing multiple index identification, so as to obtain purified iPS cell lines (TAZ-UiPSC) having a BTHS patient urine cell source. The TAZ-UiPSC can be directionally induced and differentiated into cardiac muscle cells to establish an in vitro cell model, and is used for researching of BTHS pathogenic mechanisms and screening of targeted medicines.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically refers to Bartholin's syndrome inducible pluripotent stem cells and a preparation method, differentiation medium and application thereof. Background technique [0002] In 1983, Bath Syndrome (BTHS) was first discovered by Dr. Peter Bath and is widely recognized as a rare autosomal recessive disorder linked to the X chromosome. The disease includes skeletal myopathy, cardiomyopathy, growth retardation and 3-methylglutaconic aciduria, etc., and the incidence rate is only 1 / 300000-400000. Clinical features of Barth syndrome include hypertrophic cardiomyopathy, dilated cardiomyopathy, LV noncompaction, ventricular arrhythmias, endocardial fibroelasticity, proximal myopathy, sudden cardiac death, neutropenia , lethargy and fatigue, hypoglycemia, lactic acidosis, difficulty eating, etc. [0003] In 1996, the potential mutations of Barth syndrome were mapped to the tafazzin (TA...

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Problems solved by technology

[0004] Because human cardiomyocytes are difficult to obtain, the difficulty is high and the cost is high

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