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Artificial transcription factor induced pluripotent stem cells

A cell and transcriptional regulation technology, applied in the field of pluripotent stem cells, which can solve problems such as low efficiency

Active Publication Date: 2011-09-21
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the efficiency of this method is very low, about 0.01%-0.1%

Method used

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  • Artificial transcription factor induced pluripotent stem cells
  • Artificial transcription factor induced pluripotent stem cells
  • Artificial transcription factor induced pluripotent stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0118] Materials and Methods

[0119] Cell culture: Mouse ES cells and iPS cells were maintained on a feeder layer of mitomycin C-treated mouse embryonic fibroblasts (MEFs) in DMEM (Invitrogen) supplemented with 15% heat-inactivated fetal calf Serum (FBS, Invitrogen), 2 mM L-glutamine, 0.1 mM non-essential amino acids, 1 mM sodium pyruvate, 0.1 mM β-mercaptoethanol (sigma), 1000 units / ml of leukocyte inhibitory factor (LIF, Chemicon) and 50 Units / 50mg / ml of penicillin and streptomycin. Oct4-GFP MEFs were produced from E13.5 embryos obtained from the crossing of male TgOG2 transgenic mice 22 and female wild-type C57BL mice. MEFs were grown in DMEM supplemented with 10% FBS (Hyclone), 2 mM L-glutamine, 0.1 mM non-essential amino acids, and 100 units / 100 mg / ml of penicillin and streptomycin. iPS cells were generated using MEFs of previous passages (up to passage 4).

[0120] Human ES and iPS cells were maintained with 20% Knockout Serum Replacement (KSR, Invitrogen), 2 mM L-gl...

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Abstract

The invention relates to an artificial factor and application thereof in reprogramming somatic cells into induced pluripotent stem cells and other cell lineages, in particular to a fusion protein containing a protein coded by a cell totipotency related gene and a transcriptional regulation structure domain, a coding sequence and an expression vector thereof, and a composition thereof. The invention also relates to a method for reprogramming somatic cells into induced pluripotent stem cells and other cell lineages, and cells containing the fusion protein and the coding sequence.

Description

technical field [0001] The present invention relates to the field of pluripotent stem cells. In particular, the invention relates to the use of artificial factors to reprogram somatic cells into pluripotent stem cells or other types of cells. Background technique [0002] Embryonic stem cells are derived from the inner cell mass of blastocyst embryos, capable of self-renewal and maintaining totipotency (Evans and Kaufman, 1981; Martin, 1981). In 1998, Thomson successfully established and cultivated a pluripotent human embryonic stem cell line (Thomson et al., 1998). Subsequent studies have proved that the human embryonic stem cells established by Thomson's method can infinitely self-renew in vitro and differentiate into adults at the same time. cells of almost all tissue types. Stem cells can be cultured in vitro, and they can be directional induced to differentiate into various tissue cells required, and then these differentiated cells can be introduced into disease model...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N15/85C12N5/10C12N5/07
CPCC12N2501/603C07K14/52C12N2501/602C12N2510/00C12N2501/605C12N5/0696C12N2501/604C12N2830/34C12N2501/60
Inventor 徐国良王旸
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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