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Caspase-1-ES cell line induced by mifepristone

A mifepristone and cell line technology, applied in the field of cell lines, can solve the problems of reducing ES cell transplantation into tumors, and achieve the effects of reducing the risk of tumor formation, improving safety, and maintaining curative effect.

Inactive Publication Date: 2013-12-04
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report about the transfer of caspase-1 into ES cells, which not only maintains the efficacy of ES therapy, but also reduces the risk of ES cell transplantation into tumors.

Method used

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  • Caspase-1-ES cell line induced by mifepristone
  • Caspase-1-ES cell line induced by mifepristone
  • Caspase-1-ES cell line induced by mifepristone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 The expression of caspase-1 induced by mifepristone eliminates the tumor formed by embryonic stem cells

[0054] 1. Purpose of the experiment

[0055] Caspase-1 is one of the inflammatory caspases, which can lead to inflammatory cell death after activation. Overexpression in vivo can also lead to tumor cell death. After activation, Caspase-1 can induce the formation of pores in the plasma membrane to allow water to enter the cell, causing osmotic lysis; resulting in the release of pro-inflammatory cell contents and caspase-1-related cytokines such as IL-1β and IL-18; it can also activate a certain An unknown endonuclease cleaves DNA. Therefore we chose this gene as the killer gene.

[0056] By introducing the GeneSwitch technology platform into ES cells, a caspase-1 expression system regulated by mifepristone was constructed, and the effects of high expression of caspase-1 on undifferentiated ES cells and Effects on generated tumor cells.

[0057] 2...

Embodiment 2

[0120] Example 2 Expression of caspase-1 induced by mifepristone eliminates tumors formed by embryonic stem cells in the brain but not transplanted dopaminergic neurons

[0121] 1. Purpose of the experiment

[0122] The study used the caspase-1-ES cell line regulated by mifepristone to eliminate the tumor cells formed by ES cells while retaining the dopaminergic neurons formed by differentiation of ES cells.

[0123] 2. Materials and methods

[0124] 1. Main reagents and materials

[0125] Rabbit anti-Oct-4 antibody (Cell Signaling, USA), rabbit anti-tyrosine hydroxylase (TH) antibody, mouse anti-nestin antibody (Chemicon, USA), mouse anti-SRY antibody (Abcam, UK), mouse anti-MAP2 antibody , mouse anti-microtubule-associated protein 2 (MAP 2) antibody (Sigma, USA), goat anti-MAP2 antibody (Santa Cruz, Germany), Prolong gold antifade reagent, Alpha-MEM culture medium, Alexa Fluor? 350 donkey anti-goat antibody ( Invitrogen, USA), laser confocal microscope (LCM510) (Z...

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Abstract

The invention relates to a caspase-1-ES cell line induced by mifepristone. The collection number of the caspase-1-ES cell line is CCTCCC201193. The invention also provides a preparation method and application of the cell line. The invention has the advantages that: caspase-1 is transferred into an embryonic stem (ES) cell for the first time, and the differential potential of the cell is not influenced; the caspase-1 is highly expressed in the ES cell for the first time, it is proved that activated caspase-1 initiates DNA damage to cause death of undifferentiated caspase-1-ES cells in vitro, and a risk of nodule formation caused by ES cell transplantation is reduced; it is discovered that the highly expressed caspase-1 in the differentiated ES cell does not obviously influence the cell for the first time, and the treatment effect of the ES cell is maintained; and the safety of embryonic stem cell transplantation is improved, the effect of treating parkinson disease and other neurologic diseases is remained, and the caspase-1-ES cell line can be widely applied to stem cell transplantation.

Description

technical field [0001] The invention relates to a cell line, in particular to a mifepristone-induced caspase-1-ES cell line and a preparation method and application thereof. Background technique [0002] Parkinson's disease (PD) is the second most common neurodegenerative disease, and its pathological features are mainly the loss of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies. Commonly used treatment strategies are limited to symptomatic treatment and try to alleviate the clinical symptoms of patients. Such treatment methods cannot fundamentally reverse the occurrence and development of PD. Embryonic stem cells (ES cells) are cells with multiple differentiation potentials, which can be induced to differentiate into specific cell types. This characteristic makes them have broad prospects in cell replacement therapy. ES cell transplantation is expected to be applied to the treatment of PD, but a small amount of undifferentiated ES cells mixed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/85A61K35/54A61P1/16A61P3/10A61P9/00A61P9/10A61P19/04A61P19/08A61P21/04A61P25/00A61P25/14A61P25/16A61P29/00A61P35/02A61K35/545
CPCY02A50/30
Inventor 乐卫东王颐杨德华
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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