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A skin reconstruction method based on hair follicle stem cells and silicone gel dressing

A technology of hair follicle stem cells and silicone gel dressing, applied in the biological field, can solve the problems of long surgical suture operation time, low hair follicle regeneration efficiency, poor repeatability, etc., achieve strong practical application prospects and scientific research value, strong in vitro proliferation ability, Improved reproducibility of the effect

Active Publication Date: 2016-01-27
广州汉氏联合生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there are many methods for hair follicle reconstruction experiments, these methods have more or less shortcomings such as high hardware requirements, long operation time, high difficulty, low hair follicle regeneration efficiency, and poor repeatability. The material cost is high, the surgical operation is difficult, the reconstruction area is small, and the wound exposure time is long; the subcutaneous injection (Patch) method is subcutaneous injection to achieve reconstruction, so the operation is difficult, the reconstruction area is limited, the efficiency is low, and the repeatability is poor; dissection Because the (Flap) method is a layered operation of the skin, the operation is difficult, takes a long time, the regeneration efficiency is low, and the repeatability is poor. Shortcomings impede the research and application of hair follicle stem cells in skin regeneration
Although there are commercially available artificial epidermis at home and abroad, this kind of artificial skin lacks elasticity after transplantation to the affected area due to the lack of accessory structures such as sebaceous glands and sweat glands, and it cannot perform the full function of normal skin

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  • A skin reconstruction method based on hair follicle stem cells and silicone gel dressing
  • A skin reconstruction method based on hair follicle stem cells and silicone gel dressing
  • A skin reconstruction method based on hair follicle stem cells and silicone gel dressing

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1 Preparation of Rat Prominence Hair Follicle Stem Cells

[0048] There are many sources of hair follicle stem cells required for skin reconstruction, for example, they can be obtained by means of flow cytometry according to marker molecules of hair follicle stem cells, or obtained by tissue and organ culture.

[0049] In this example, taking adult rat vibrissae hair follicle stem cells as an example, the method for preparing seed cells for hair follicle reconstruction of the present invention is described in detail, as follows. 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.

[0050] 1. Microscopic stripping of rat vibrissae hair follicles: take a small piece of adult Wistar rat (Institute of Zoology, Chinese Academy of Sciences) vibrissae skin, disinfect with 75% alcohol for 3-5 minutes; under a stereoscope, cla...

Embodiment 2

[0054] Example 2 Fluorescence labeling of stem cells

[0055] 1. Preparation of hair follicle stem cells: resuscitate adult rat vibrissae hair follicle stem cells prepared and frozen in Example 1, inoculate a small amount of fibroblasts as trophoblast cells according to the established hair follicle stem cell culture system, and use William's E complete medium in 37°C, 5% CO 2 , 100% humidity incubator.

[0056] 2. Hair follicle stem cells were labeled with green fluorescent protein: after 24 hours of restorative growth, the cells were in good growth state; add the lentiviral stock solution (purchased from Invitrogen) with the green fluorescent protein gene, and replace it with fresh William 'sE complete medium, green cells can be observed in 48 hours.

[0057] 3. Purification and expansion of green fluorescent protein-labeled hair follicle stem cells: first, using the characteristics of hair follicle stem cells to adhere tightly to the wall, use digestive solution (0.1% t...

Embodiment 3

[0058] The preparation of embodiment 3 neonatal rat dermal cells

[0059] Rats within 24 hours after birth were killed in ice, soaked in 75% alcohol for 3-5 minutes, and then operated in an ultra-clean workbench. First, they were washed twice with sterile saline. The skin on the entire back was cut off with tweezers, washed once with normal saline, then the subcutaneous fat was gently scraped off with a scalpel, and then washed seven times with normal saline. Spread the skin epidermis up in a Petri dish, carefully add digestive solution (0.25% trypsin and Dispase (Gibco) at a volume ratio of 1:1), float the skin on the liquid surface, and digest overnight at 4°C. The epidermis was peeled off the next day, and the dermis was digested in type IV collagenase at 37°C for 45min. Filter the digested dermal cell suspension with a 400-mesh sieve, collect the filtered single-cell suspension, centrifuge at 1000 rpm for 10 min to collect the cells, count them, and resuspend them in DM...

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Abstract

The invention relates to a skin renewal method. According to the skin renewal method, hair follicle stem cells are used as seed cells and silica gel dressing is used as a new cell transplantation carrier. The skin renewal method comprises the following steps: 1) cultivating and amplifying hair follicle stem cells; 2) cultivating dermal cells; 3) mixing the hair follicle stem cells cultivated and amplified in the step 1) and the dermal cells cultivated in the step 2); 4) dropwise adding mixed cells obtained in the step 3) into silica gel dressing for incubation; and 5) transplanting the cells and the silica gel dressing used in the step 4) to a part required to be subjected to skin renewal. The method provided by the invention not only can renew functional skin in the treatment of large-area skin injuries, but also can provide a simple validation method for the multipotency test on hair follicle stem cells in scientific research activities.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel skin reconstruction method based on hair follicle stem cells and silicone gel dressing. Background technique [0002] The skin is the largest organ of mammals. It can not only act as a barrier to protect the body from harmful factors such as external pathogens, but also contains abundant nerve endings that can quickly sense environmental changes. In addition, the skin also plays an important role in temperature regulation and water retention. also play an irreplaceable role. However, when faced with problems such as skin damage caused by burns or infectious diseases, the repair and regeneration of skin has always been a major problem that plagues human beings. [0003] As an important part of the skin, the hair follicle also plays an important role in many aspects such as protecting the body surface, sensory conduction, hair color and skin color determination, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074A61L27/38A61L27/60
Inventor 段恩奎赵华山张会闪张守兵乔静巧
Owner 广州汉氏联合生物科技有限公司
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