Sub totipotential stem cell and preparation method and application thereof

A technology of sub-totipotent stem cells and uses, applied in the field of sub-totipotent stem cells, the preparation of sub-totipotent stem cells, can solve the problems of poor understanding of stem cells

Active Publication Date: 2010-06-23
BEIJING HEALTH & BIOTECH (H&B) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, people's understanding and clinical use of multipotent stem cells such as hematopoietic stem cells is relatively deep, and the understanding of embryonic stem cells has also made great progress. Many characteristics of stem cells, but also have many biological characteristics of pluripotent stem cells, transplanted into the body will not produce teratomas, such a type of stem cells are poorly understood

Method used

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  • Sub totipotential stem cell and preparation method and application thereof
  • Sub totipotential stem cell and preparation method and application thereof
  • Sub totipotential stem cell and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Isolation of CD151 by adherent subculture + CD184 - Oct4 + subtotipotent stem cells

[0073] Take cut human placenta, carry out CD151 + CD184 - Oct4 + Preparation of stem cell products. The specific steps are as follows: 1) Aseptically collect the cut human placenta; 2) Cut to small fragments of 3 Small tissue pieces were digested with 0.5mg / mL collagenase I at 37°C for 1 hour, and 1mg / mL trypsin at 37°C for 30 minutes, with gentle shaking 5 times during the period, sieved, centrifuged to collect the cell suspension, and inoculated in T75cm 2 In the culture flask, cultured in cell culture solution A for 5 to 7 days, the spindle-like cells adhered to the wall; 3) The adherent cells were digested with trypsin and further cultured, passaged, purified, expanded and passaged for 6 generations, and the cells were harvested, After concentrating and preparing a cell suspension with cell preservation solution, some cells are sent for quality inspection, and the rest are st...

Embodiment 2

[0077] Isolation of CD151 by adherent subculture + CD184 - Oct4 + subtotipotent stem cells

[0078] Same as Example 1, except that the cell culture medium used is cell culture medium B.

[0079] The results obtained were substantially the same as in Example 1.

Embodiment 3

[0081] Isolation of CD151 by adherent subculture + CD184 - Oct4 + subtotipotent stem cells

[0082] Same as Example 1, except that the cell culture medium used is cell culture medium C.

[0083] The results obtained were substantially the same as in Example 1.

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Abstract

The invention discloses a method for preparing a population of?human pluripotent stem cells and the application thereof. The preparation of stem cells is characterized by comprising the following steps: CD151<+>, CD31<->, Sox<2+> pluripotent stem cells are separated and collected from human umbilical cord and or placenta tissues; the cells adhere to grow in a culture vessel under a predetermined condition and expand through passage 20 or above to be still stable in gene expression. The population of cells of this invention do not form teratoma after injection into animals. The human pluripotent stem cells highly express CD151, OCT4 and Sox-2 as specific markers of embryonic stem cells, as well as specific markers of epidermic cells, endothelial cells, thrombocytes, dendritic cells, while lack expression of CD31, CD34, CD45 and HLA-II. The pluripotent stem cells are also characterized as being able to adhere to tissue culture plastic and having the potential to differentiate into three germ layers: endoderm, mesoderm and ectoderm. These pluripotent stem cells are able to be used as carrier cells of gene therapy and for the treatment of diseases caused by cell damage or cell aging. The present invention provides a method of isolating, purifying and culturally expanding of a population of human pluripotent stem cell for preparing the high purity injection preparation. The preparation of stem cells has a good therapeutic effect on the treatment of diseases caused by cell damage or cell aging in animal and human clinical trials. The preparation also has no toxic side effect and no immune rejection.

Description

technical field [0001] The present invention relates to the field of stem cells, in particular to a subtotipotent stem cell derived from cut human umbilical cord or placenta. At the same time, the present invention also relates to the preparation method of the sub-totipotent stem cells and the use of the sub-totipotent stem cells. Background technique [0002] The human body is made up of trillions of cells. Cells are the basic structural and functional units of the human body, which are differentiated from stem cells, so stem cells are the origin cells of various tissues and organs in the human body. Stem cells are characterized by self-replication and multilineage differentiation. During normal reproductive physiology, stem cells are differentiated from fertilized eggs. The fertilized egg initially develops into primitive embryonic stem cells, which have the full potential to form a complete individual. Primitive embryonic stem cells further differentiate and prolifera...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A61K48/00A61K35/48A61K35/44A61P7/06A61P37/02A61P25/00A61P37/06A61P39/06A61K35/12
CPCC12N2500/25C12N2501/115C12N2501/11A61K2035/122C12N2501/39A61K35/12C12N5/0607C12N2500/36C12N5/0605A61P7/06A61P25/00A61P37/00A61P37/02A61P37/06A61P39/06
Inventor 赵宝娜韩之海
Owner BEIJING HEALTH & BIOTECH (H&B) CO LTD
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