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A method for culturing induced pluripotent stem cells using human urine cells as a feeder layer

A technology of pluripotent stem cells and feeder layer cells, which is applied in the field of culturing induced pluripotent stem cells, can solve the problems of difficulty in obtaining cells and heterologous contamination of human induced pluripotent stem cells, and achieve the effect of good growth state.

Inactive Publication Date: 2021-02-19
GUANGDONG XTEM BIOTECH CO LTD
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Problems solved by technology

But the mouse embryonic fibroblasts (MouseEmbryonic Fibroblasts, MEF) used in the traditional method will bring heterogeneous contamination to the cultured human induced pluripotent stem cells. "Methods for Sexual Pluripotent Stem Cells" (CN 102161980A), using bone marrow mesenchymal stem cells as a feeder layer, there is a limitation that the cells are difficult to obtain

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  • A method for culturing induced pluripotent stem cells using human urine cells as a feeder layer
  • A method for culturing induced pluripotent stem cells using human urine cells as a feeder layer
  • A method for culturing induced pluripotent stem cells using human urine cells as a feeder layer

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[0016]Hereinafter, the present invention will be further described with reference to the drawings and specific embodiments to better understand the present invention. The materials and reagents used in the examples can be obtained from commercial sources unless otherwise specified. Example Urine cells are used as feeder layers to cultivate human induced pluripotent stem cells

[0017]The present invention provides a method for culturing human induced pluripotent stem cells by using human urine cells to make feeder layers. For details, seefigure 1 .

[0018]1. Separation and culture, cryopreservation and recovery of urine cells

[0019]1) Separation and culture of urine cells. Transfer fresh urine cells to a 50mL centrifuge tube with 5mL double antibody (operate under aseptic conditions to avoid contamination), centrifuge at 400g for 10 minutes, discard the supernatant, and leave 1mL on top Supernatant, resuspend cells, transfer all cells to the same 50mL centrifuge tube, add 10mL PBS to wash...

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Abstract

The invention provides a method for cultivating human induced pluripotent stem cells by using human urine cells as a feeder layer. Human pluripotent stem cells (hiPS) reprogrammed from adult male foreskin fibroblasts were cultured with subcultured human urine cells from the third to twelfth passages cultured after cryopreservation and thawing as trophoblast cells. The human urine cells used in the present invention replace mouse embryonic fibroblasts or mouse embryonic fibroblast cell lines used in traditional methods to cultivate human induced pluripotent stem cells, which reduces the contamination of heterologous cells in the culture system, and is more effective than mesenchymal Mesenchymal stem cells are easier to obtain. And it was proved that this culture system can expand human induced pluripotent stem cells in vitro, and maintain the biological characteristics and pluripotent stemness of hiPS cells for a long time, which provides the possibility for hiPS to enter clinical application.

Description

Technical field[0001]The present invention relates to the field of biotechnology, in particular to a method for culturing induced pluripotent stem cells using human urine cells as feeder layers.Background technique[0002]Induced pluripotent stem cell technology is considered to be the most remarkable stem cell technology since the 21st century. In August 2006, Takahashi and others of Kyoto University, Japan[1]It was announced on "Cell" that they selected four genes (Oct4, Sox2, c-Myc, and Klf4) from the 24 genes related to the pluripotency of embryonic stem cells, and introduced the four selected genes into mice. After skin fibroblasts, mouse skin fibroblasts are reprogrammed into a new type of cells with characteristics similar to embryonic stem cells, which can express embryonic stem cell-specific surface markers and have the ability to differentiate into various types of cells of the three germ layers The potential is called induced pluripotency of the cell. The acquisition of hum...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/867
CPCC12N5/0696C12N15/86C12N2501/602C12N2501/603C12N2501/604C12N2501/606C12N2502/1323C12N2510/00C12N2740/10043
Inventor 于云飞陈勇彭特蔡亚雄刘樱乔志平
Owner GUANGDONG XTEM BIOTECH CO LTD
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