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Method for preparing induced pluripotent stem cells and its applications

A technology of pluripotent stem cells and cells, applied in the field of preparation of induced pluripotent stem cells, can solve the problem of unknown protective role mechanism of iPSCs, etc.

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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the possible protective roles of iPSCs and the underlying mechanisms remain unknown

Method used

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  • Method for preparing induced pluripotent stem cells and its applications
  • Method for preparing induced pluripotent stem cells and its applications
  • Method for preparing induced pluripotent stem cells and its applications

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Experimental program
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Embodiment -(I

[0068] I. Materials and methods

[0069] 1. Generation and cell culture of iPSC lines

[0070] C57 / B6 mice were used in this study and all procedures involving animals were approved by the Animal Committee of Taipei Veterans General Hospital. Murine iPSCs are produced from mouse embryonic fibroblasts (MEFs) derived from 13.5-day-old C57 / B6 mouse embryos. The iPSCs were reprogrammed by transduction of retroviral vectors with four transcription factors (Oct4 / Sox2 / Klf4 / c-Myc; OSKM) or three transcription factors (Oct4 / Sox2 / Klf4; OSK) encoding, as previously described (Li et al., 2011). A total of 12 strains (Re-1 to Re-12; OSKM) were selected and established. Re-7 iPSCs strains have passed through more than 100th passage stably and have high pluripotency. Therefore, Re-7 iPSCs were selected and widely used in this study. Briefly, undifferentiated iPSCs were routinely cultured and expanded in six-well culture dishes on mitosis-inactivated MEFs (50,000 cells / cm 2 ) (BD Techno...

Embodiment -(II

[0115] I. Ventilator-Induced Lung Injury Model

[0116] Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are diseases of acute respiratory failure and apparently non-cardiogenic pulmonary edema, respiratory distress and hypoxemia. High tidal volume-induced mechanical ventilation in patients has been shown to increase the risk of pathological hyperinflation in the lungs, eliciting production of inflammatory mediators, new inflammatory cells and ultimately induction of a type of ALI called ventilator-induced Ventilator-induced lung injury (VILI). Recently, the therapeutic efficacy of mesenchymal stem cells (MSCs) in modulating inflammatory responses has been demonstrated in sepsis-induced ALI (Mei SH et al., Am J Respir Crit Care Med 2010; 182:1047-1057; with Chien MH et al. , Crit Care Med 2012;40:1245-1253). Another study further indicated that MSC therapy enhanced lung repair in VILI through a keratinocyte growth factor-dependent paracrine mechanism (C...

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Abstract

The present invention relates to a novel method for preparing induced pluripotent stem cells (iPSCs) by introducing three genes, Oct3 / 4, Sox2, and Parp1, into somatic cells. The present invention also relates to the iPSCs produced by the aforementioned method. Also provided is a method of rejuvenating cells by use of a PARylated protein or an enzyme with PARylation activity. Further provided is a method for inducing the secretion of interferon-γ inducible protein-10 (IP-10) comprising administering to a subject in need thereof an effective amount of iPSCs or iPSC-CM.

Description

[0001] Cross-Referenced Related Applications [0002] This patent application claims priority to US Provisional Patent Application Serial No. 61 / 708,128, filed October 1, 2012, which is hereby incorporated by reference in its entirety. This application also claims priority to US Provisional Patent Application Serial No. 61 / 717,871, filed October 24, 2012, which is hereby incorporated by reference in its entirety. technical field [0003] The present invention generally relates to a method for preparing induced pluripotent stem cells, and a method for rejuvenating cells through the method. The invention also relates to a method for treating tissue damage. Background technique [0004] Cell reprogramming techniques for the generation of induced pluripotent stem cells (iPSCs) are promising strategies for stem cell biology and regenerative medicine. Previous data have shown that nuclear reprogramming can be experimentally induced by three methods: nuclear transfer, cell fusion...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N5/10A61K35/545
CPCC12Y204/0203A61K35/12A61K38/46A61K38/45C12Y306/04012A61K35/545A61P1/16A61P11/00
Inventor 邱士华简越邱光裕杨逸萍
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