Stem cell repairing material as well as preparation method and application thereof

A repair material and stem cell technology, applied in the field of tissue engineering, can solve problems such as wrinkle recurrence, achieve the effect of less trauma and pain, avoid immune rejection, and avoid ethical disputes

Inactive Publication Date: 2011-10-05
王影
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since these fillers are biodegradable materials, wrinkles will reappear in the short term

Method used

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  • Stem cell repairing material as well as preparation method and application thereof
  • Stem cell repairing material as well as preparation method and application thereof
  • Stem cell repairing material as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Extraction, cultivation and identification of adipose-derived mesenchymal stem cells

[0032] 1. Extraction and culture of adipose-derived mesenchymal stem cells

[0033] 1) After removing excess components such as blood vessels and connective tissue in 30 ml of adipose tissue obtained by instrumented liposuction, rinse with 0.8% injectable saline containing antibiotics for 2-3 times to obtain light yellow fat granule tissue;

[0034] 2) Add 2 mg / ml type I collagenase solution (Worthington, USA) to the purified fat particles, digest at 37°C for 60-90 min, and stop with complete medium (DMEM+10% FBS, both from Gibco, USA) Digestion;

[0035] 3) 150-mesh sieve to filter the digested product of human adipose tissue;

[0036] 4) After filtration, centrifuge at 1000 rpm for 10 min, resuspend in complete medium, stain with Trypan blue (Sinopharm Group) and count;

[0037] 5) In complete medium at 1×10 5 / cm 2 Human adipose-derived mesenchymal stem cells were in...

Embodiment 2

[0046] Example 2 Preparation and detection of adipose-derived mesenchymal stem cells / autologous serum complex

[0047] 1. Preparation of Adipose-derived Mesenchymal Stem Cell / Autologous Serum Complex

[0048] According to the method described in Example 1, the extraction, isolation, culture and expansion of adipose-derived mesenchymal stem cells were carried out, and the obtained adipose-derived mesenchymal stem cells were mixed with autologous serum to prepare a cell concentration of 5×10 6 / ml of cell suspension. This suspension can be directly applied to clinical treatment.

[0049] 2. Detection of Adipose-derived Mesenchymal Stem Cell / Autologous Serum Complex

[0050] (1) Detection of cell survival rate: use trypan blue exclusion test to measure the cell survival rate and cell number of the collected cells, and the cell survival rate is above 95%;

[0051] (2) Detection of cell sterility and virus-free: refer to the third appendix of "Chinese Pharmacopoeia" 2005 edition...

Embodiment 3

[0053] Embodiment 3 Animal experiments

[0054] The cell / serum complex in Example 2 was injected subcutaneously into nude mice via a 1 ml syringe. Before the injection, the nude mice were anesthetized, and the injection volume at each injection point was controlled at about 250ul. The control group used an equal volume of normal saline instead of the injection. Five nude mice (weight about 20 g, healthy, male) were selected for the experimental group and the control group each time, and the experiments of the two groups were repeated three times under the same conditions. The nude mice in the experimental group and the control group were dissected and observed 30 days after the injection experiment, and the skin at the injection site was collected for tissue section staining. The results showed that exogenous human adipose stem cells existed in the skin of the injection site of the nude mice in the experimental group, and the keratin antibody was positive. In vivo differenti...

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Abstract

The invention relates to a stem cell repairing material as well as a preparation method and application thereof. The stem cell repairing material comprises a stem cell and a cell carrier, wherein the stem cell is an autologous adipose-derived mesenchymal stem cell (ADSC) with a concentration of 105-108/ml, and the cell carrier is autoserum, normal saline (0.9%) or a glucose injection (5%). In the invention, adopted seed cells are prepared through separating and purifying autologous adipose tissues of patients, thereby avoiding the ethical disputes and the immunological rejection. The material drawing of the adipose tissues can be performed by using an instrument suction method, which is simple in operation and can bring less traumas and pains to the patients. The cell carrier used in the invention is the autoserum, normal saline (0.9%) or glucose injection (5%), therefore, the cell carrier mixed with mesenchymal stem cells and other ingredients can be injected into the patents by virtue of syringes. By using the stem cell repairing material provided by the invention, the operation process is simple, no scar is left, and the wounds and pains for the patients are avoided, therefore, the stem cell repairing material can be easily accepted by the patients.

Description

technical field [0001] The invention relates to a material suitable for plastic surgery and cosmetic surgery, in particular to an injectable stem cell repair material, its preparation method and application, belonging to the field of tissue engineering. Background technique [0002] Anti-aging is a topic of common concern to all mankind. Most of the substances that support human skin exist in the dermis. A fiber network composed of collagen, elastic fibers and hyaluronic acid supports the skin, and its content determines the texture of the skin. All three substances are secreted by fibroblasts in the dermis: fibroblasts constantly secrete collagen, elastic fibers and hyaluronic acid to maintain the luster and texture of the skin. After the age of 25, fibroblasts decrease at a rate of 1% to 2% per year, followed by a decrease in the number of collagen, elastic fibers and hyaluronic acid, showing signs of aging. The attenuation of fibroblasts is the main cause of skin aging....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61K35/12A61P17/02A61K35/28
Inventor 王影徐妍魏翠黄金龙
Owner 王影
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