Structural method and application of tissue engineering adipose tissue

A technology of adipose tissue and tissue engineering, applied in biochemical equipment and methods, tissue culture, bone/connective tissue cells, etc., can solve problems such as loss of cell activity, and achieve broad application prospects and huge market value potential

Active Publication Date: 2007-02-14
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is because 80% to 90% of the cytoplasm of mature adipocytes is composed of lipid droplets, which are susceptible to mechanical damage and l

Method used

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  • Structural method and application of tissue engineering adipose tissue
  • Structural method and application of tissue engineering adipose tissue
  • Structural method and application of tissue engineering adipose tissue

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0050] Example 1 In vitro isolation and culture expansion of adipose tissue-derived stem cells:

[0051] Rinse the aspirate obtained from liposuction repeatedly with phosphate buffered saline solution (PBS), then add PBS to it, stir and place until clarified to remove damaged tissue, blood and some red blood cells, etc., and then add collagen with a concentration of about 0.1% Enzymes, digested at 37°C for 45 minutes with gentle agitation. Centrifuge at 1500 rpm for 5 minutes, collect the cell pellet, and suspend the pellet in erythrocyte lysate (Tris-NH 4 C1), after standing at room temperature for 10 minutes, the cells were collected by centrifugation again. This process was repeated twice to break up and remove as many red blood cells as possible. The cell suspension was centrifuged to collect the lower cell sediment, and then the viability of the cells was detected by the bromophenol blue dye exclusion method and the proportion of living cells was counted.

[0052] Surv...

Example Embodiment

[0053] Example 2 Identification of surface markers of adipose tissue-derived stem cells

[0054] The stem cells derived from adipose tissue cultured and expanded at passage 5 were collected, digested with 0.25% trypsin, and centrifuged. After washing the cells twice with PBS, resuspend the cells with 1ml PBS and count. Adjust the cell concentration to 1 x 10 with PBS 6 pcs / ml, according to 5×10 5 Each cell / tube was divided into several EP tubes, and PE or FITC-labeled multiple antibodies (CD90, CD44, CD45, CD71, CD16, CD30, CD34, CD29, CD3, CD11b, CD133 and CD19) were tested separately. Combine, add to each tube, incubate at room temperature for 30min, then wash twice with PBS, add 0.5ml PBS to resuspend the cells, and use flow cytometry to detect and identify the surface markers of the cultured and amplified cells. The results showed that the adipose tissue-derived stem cells had positive expression of CD90, CD29 and CD71, indicating that they were a type of stem cells der...

Example Embodiment

[0055] Example 3 Directed Induced Differentiation and Application of Adipose Tissue-derived Stem Cells to Adipocytes

[0056] Use DMEM medium containing 10% FBS, 1 μM dexamethasone, 10 μM insulin, 200 μM indomethacin, 1% penicillin / streptomycin, 0.5 mM IBMX under normal conditions (37 ° C, 5% CO 2 ) to continue culturing the adipose tissue-derived stem cells. After culturing for 7-15 days, take the induced cultured cell clones, fix with methanol for 2 minutes, rinse with 50% alcohol, add Oil Red O dye for 10 minutes, rinse with 50% alcohol, rinse with water, counterstain with hematoxylin for 1 minute, observe the staining under a microscope In normal cases, oil red O staining was positive, which indicated the presence of lipid vesicles in the cells. According to this feature, it can be judged that the stem cells have shown a tendency to differentiate into mature adipocytes. Adipose tissue-derived stem cells that are undifferentiated or partially differentiated into adipocytes...

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Abstract

The invention relates to a method used to process directional inducing differentiation for the adipose tissue stem cell from vitro to adipose and endothelial cell, combine the processed the cell with biomaterial to form engineering adipose tissue which belongs to biomedicine field. It produced adipose tissue can be used to remedy soft tissue defect, repair wound tissue, and fill in normal tissue, such as face and neck tumor cutting, various complex wound, congenital malformation repair, etc. Thus it has huge market value potential and wide application prospect.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to inducing differentiation of stem cells from adipose tissue to fat and endothelial cells in vitro, and combining the differentiated cells with biological materials to jointly construct tissue engineered adipose tissue The method can be used to prepare filling plastics for soft tissue defects, wound tissue repair and normal tissue. Background technique [0002] According to the latest data released by the World Health Organization, the incidence of breast cancer in the world has almost doubled in the past 20 years, and there are currently about 1 million breast cancer patients in the world. Breast cancer has become the number one health killer of women after skin cancer. In China, the incidence of female breast cancer is also gradually increasing, from 17 per 100,000 in 1999 to 52 per 100,000 in 2004, an increase of more than three times. Among them, the highest incidence rate ...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/06C12N5/077
Inventor 裴雪涛管利东王韫芳闫舫李绍青白慈贤岳慧敏南雪
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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