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Method of generating induced pluripotent stem cells and differentiated cells

A technology for pluripotent stem cells and differentiated cells, which is applied in the field of generating induced pluripotent stem cells and differentiated cells, and can solve the problems of low efficiency and unsatisfactory clinical application.

Inactive Publication Date: 2018-04-10
UNIVERSITÄT FUR BODENKULTURE +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Since these methods are minimally invasive, require small amounts of blood, and do not require prolonged cell culture, they represent a significant advance despite the fact that they are very inefficient
However, the primary donor cells used according to these reports are mature T cells carrying specific T cell receptor rearrangements, which is undesirable for some potential clinical applications

Method used

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  • Method of generating induced pluripotent stem cells and differentiated cells
  • Method of generating induced pluripotent stem cells and differentiated cells
  • Method of generating induced pluripotent stem cells and differentiated cells

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Embodiment Construction

[0093] Materials and methods

[0094] immunofluorescence microscopy

[0095] Cells were fixed overnight in 4% paraformaldehyde, washed, blocked and permeabilized in blocking solution (PBS containing 3% normal goat serum and 0.2% Triton X-100) for 30 minutes. They were then incubated overnight at 4°C with primary antibodies in blocking solution, washed twice, and incubated with corresponding secondary antibodies for 1 hour at room temperature. Cells were washed twice and stained with DAPI (Sigma) for 5 minutes, then observed and photographed with a Leica TCS SP2 spectral confocal microscope (Leica Microsystems GmbH, Wetzlar, Germany). Prior to immunofluorescence, beating areas were cut with scissors, collected in 1.5 mL tubes with low calcium PBS, and left at room temperature for 30 minutes. These cell clumps were transferred to enzyme buffer containing 0.5-1 mg / mL collagenase 2 and incubated at 37° for 30-40 minutes. Digestion was terminated with DMEM / Ham's F121:1 (Hyclone)...

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Abstract

Methods for generating iPSCsand differentiated cells of interest by reprogramming donor cells that have been obtained in a non-invasive manner. In particular, the donor cells are exfoliated epithelialurine cells. The differentiated cells can be obtained by differentiation of the reprogrammed iPSCsor by direct reprogramming the urine cells.

Description

[0001] This application is a divisional application of the Chinese invention patent application with application number 201180063411.X, application date 2011.12.23, and invention title "Method for Producing Induced Pluripotent Stem Cells and Differentiated Cells". [0002] The present invention relates to induced pluripotent stem cells and differentiated cells and methods of producing them. Background of the invention [0003] Embryonic stem cells (ESCs) are cells derived from blastocysts obtained by in vitro fertilization that have the ability to self-renew and differentiate into any mature mammalian, eg human, cell type (a property known as "pluripotency"). Under specific tissue culture conditions, ESCs can remain undifferentiated for extended periods of time without losing their pluripotent characteristics. Owing to these properties, ESCs have attracted great interest in the field of regenerative medicine. Potentially, tissues derived from ESCs can be used for clinical tre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/0789
CPCC12N5/0647C12N5/0696C12N2506/25
Inventor 米格尔·埃斯特万约翰内斯·格里亚里雷吉娜·格里亚里裴端卿周厅
Owner UNIVERSITÄT FUR BODENKULTURE
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