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Method for culturing induced pluripotent stem cells by using human urine cells as feed layer

A technology of pluripotent stem cells and feeder cells, which is applied in the field of culturing induced pluripotent stem cells, can solve the problems of difficult to obtain cells, heterologous contamination of human induced pluripotent stem cells, etc., and achieve the effect of good growth status

Inactive Publication Date: 2018-04-20
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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  • Method for culturing induced pluripotent stem cells by using human urine cells as feed layer
  • Method for culturing induced pluripotent stem cells by using human urine cells as feed layer
  • Method for culturing induced pluripotent stem cells by using human urine cells as feed layer

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[0016] The present invention will be further described below in conjunction with specific embodiments according to the accompanying drawings, so as 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 Used as Feeder Layer to Cultivate Human Induced Pluripotent Stem Cells

[0017] The present invention provides a method for cultivating human induced pluripotent stem cells by using human urine cells as a feeder layer. For details, see figure 1 .

[0018] 1. Isolation, 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 sterile conditions to avoid contamination), centrifuge at 400g for 10 minutes, discard the supernatant, and leave 1mL Supernatant, resuspend cells, transfer all cells to the same 50mL centrifug...

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Abstract

The invention provides a method for culturing human induced pluripotent stem cells by using human urine cells as a feed layer. Third-twelfth generations of human urine cells cultured after subculture,cryopreservation and resuscitation are used as trophoblast cells for culturing the human induced pluripotent stem (hiPS) cells reassembled by adult male foreskin fibroblasts. The used human urine cells replace small mouse embryo fibroblasts or mouse embryo fibroblast cell lines used in traditional methods, the human induced pluripotent stem cells are cultured, pollution of heterologous cells in the culture system is reduced, and the human induced pluripotent stem cells are more easily obtained than mesenchymal stem cells. The culture system is proved to make the human induced pluripotent stemcells amplified in vitro, the biological characteristics and the pluripotent property of the hiPS cells are maintained for a long time, and the possibility is provided for the hiPS to enter the clinical application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for cultivating induced pluripotent stem cells using human urine cells as a feeder layer. 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 et al. from Kyoto University, Japan [1] It was announced in "Cell" that they screened four genes (Oct4, Sox2, c-Myc and Klf4) from 24 genes related to the pluripotency of embryonic stem cells, and introduced the four genes into the mouse After skin fibroblasts, mouse skin fibroblasts were 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 potential, known as induced pluripotency of the cells. The acquisition of h...

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Application Information

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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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