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Hair follicle stem cell separation culture method

A hair follicle stem cell, isolation and culture technology, applied in the direction of animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of hair follicle stem cells not easy to survive, cells are prone to apoptosis, and the efficiency of clone formation is low, so as to achieve the efficiency of clone formation High, maintaining the same characteristics, the effect of small cell damage

Active Publication Date: 2014-01-29
广州汉氏联合生物科技有限公司
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Problems solved by technology

However, this flow cytometry-based sorting method has a great disadvantage, that is, a large amount of skin is required to prepare a single-cell suspension, which is impossible for patients with extensive skin damage (burns or scalds, etc.)
In addition, during the sorting process, single cells spend a long time in vitro and are in a suspended state, so the cells are prone to apoptosis and mechanical damage when the cells pass through the flow cytometer. extremely inefficient

Method used

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  • Hair follicle stem cell separation culture method
  • Hair follicle stem cell separation culture method
  • Hair follicle stem cell separation culture method

Examples

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Embodiment 1

[0037] Example 1 : Isolation of Hair Follicle Stem Cells from a Single Rat Vibrissae Using Organ Culture

[0038] This embodiment takes a single rat tentacles as an example to describe in detail the method for isolating hair follicle stem cells provided by the present invention. For the specific technical route, see figure 1 . The feeding and use of experimental animals were carried out in accordance with the regulations of the Animal and Medical Ethics Committee of the Institute of Zoology, Chinese Academy of Sciences.

[0039] 1. Microdissection of rat vibrissae hair follicles

[0040] Take a small piece of adult Wistar rat (Institute of Zoology, Chinese Academy of Sciences) tentacles skin, disinfect with 75% alcohol for 3-5 minutes; under the stereoscope, use tweezers to clamp the isthmus of the tentacles hair follicles near the epidermis, and apply force toward the dermis Tear out the hair follicles; fix the hair follicles with a 29G needle, carefully scratch the derma...

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Abstract

The invention provides a hair follicle stem cell separation culture method. The method utilizes an organ culture method, adopts the William's E complete medium to culture the hair follicle, separate and purify to get hair follicle stem cells. The method only uses very small skin materials, the rat whisker follicle is separated with a stereoscope through a micro separation method, then the whole whisker follicle is cultured, the hair follicle stem cells grow out from the projection part and form clone, so the cell clone forming efficiency is high, the purification is easy, long-term passage can be realized, the characteristics of the hair follicle stem cells are kept unchanged, and only one whisker follicle can obtain enough hair follicle stem cells for long-term use.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating and culturing hair follicle stem cells. Background technique [0002] The development and progress of stem cell biology research has greatly promoted people's understanding of many basic biological issues in the development process, and at the same time brought people a new idea of ​​disease treatment-stem cell transplantation. For the treatment of large area skin burns (scalds) and skin diseases, stem cell transplantation has a long-term application prospect. Epidermal stem cell transplantation has been used clinically as early as 1991. However, the skin formed by it does not have sebaceous glands and hair follicles, so it is functionally incomplete. [0003] The skin is a barrier between the mammal itself and the external environment, and has physiological functions such as resisting microbial infection, maintaining moisture, regulating body tem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 段恩奎张守兵
Owner 广州汉氏联合生物科技有限公司
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