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Marker membrane protein for purifying and enriching intradermal multipotentiality cells

A technology of differentiation potential and membrane proteins, applied in the field of marker membrane proteins, can solve problems such as huge data volume and achieve the effect of convenient sorting

Inactive Publication Date: 2010-06-30
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, a huge amount of data is often obtained through these technical analysis; therefore, finding a marker (Marker) that can effectively screen MDFs in the dermis is the main problem faced

Method used

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  • Marker membrane protein for purifying and enriching intradermal multipotentiality cells
  • Marker membrane protein for purifying and enriching intradermal multipotentiality cells
  • Marker membrane protein for purifying and enriching intradermal multipotentiality cells

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Embodiment Construction

[0026] Below respectively in conjunction with accompanying drawing and embodiment the embodiment of the present invention is described further:

[0027] 1. In vitro culture and expansion of MDFs single cell clones

[0028] Human dermis-derived cells were cultured and expanded in vitro; when the cells grew to 80% confluent, they were digested with 0.25% trypsin and passed on; Dilute by limiting dilution method, add a single cell into a 96-well plate for culture, and replace the conditioned medium every 3 days until a larger clone is formed. A certain number of monoclonal cells are obtained by further subculturing the monoclonal cells (the first 4 generations).

[0029] 2. Osteogenic-fatty-nerve three-way induction and identification of MDFs

[0030] From the human MDFs monoclonal cells obtained in the above steps, 12 single-cell clones were randomly selected (one of the single-cell clones was contaminated during the passage process), and were properly passaged and amplified i...

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Abstract

The invention relates to marker membrane protein for purifying and enriching intradermal multipotentiality cells. The protein is obtained by adopting the following steps of: in vitro culturing and amplifying cells from dermis, and diluting and culturing by using a 96 orifice plate to obtain a plurality of monoclonal cells; carrying out in vitro induction of forming fat, forming bones and forming nerves on the monoclonal cells and identifying the differentiation capacity of the cells; respectively separating and purifying two-way cloned cells and three-way cloned cells to obtain corresponding membrane protein components; finding out difference protein spots of three-way clone and two-way clone through a two-way electrophoretic analysis; carrying out LTQ mass spectrum analysis to obtain matched protein; and finding out the marker membrane protein through Western-Blot detection. The invention also relates to an application of the marker membrane protein. The marker membrane protein which is primarily screened by adopting the method and presents more obvious up-regulation or down-regulation along with the change of the differentiation capacity can be used for separating and purifying cells with multipotentiality.

Description

technical field [0001] The invention relates to a marker membrane protein, in particular to the marker membrane protein used for purifying and enriching multi-lineage differentiation potential cells in the dermis. Background technique [0002] The wide application prospect of stem cells in cell therapy, gene therapy and tissue engineering has made it a research hotspot in the field of life sciences in recent years. Adult stem cells are the focus of research in this field because they do not involve medical ethics issues, and have the advantages of multi-directional differentiation potential and autologous materials. At present, adult stem cells with different differentiation abilities have been isolated from bone marrow, fat, muscle and other tissues, and some cells (such as bone marrow stromal stem cells) have been successfully used in the repair of tissue and organ defects. [0003] Skin is the organ with the largest surface area in the human body, rich in cell resources,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N9/90C12N5/071
Inventor 陈付国毕丹曹谊林张文杰刘伟崔磊
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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