Vascularized fat depot based on partition and construction method thereof

A technology of adipose tissue and construction method, applied in the field of bioengineering, can solve problems such as no breakthrough progress, difficulty in stabilization, hard breast tissue of synthetic polymer materials, etc.

Active Publication Date: 2009-07-29
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there are problems: on the one hand, the commonly used synthetic polymer materials are too hard compared to breast tissue; on the other hand, the degradable polymer materials of composite cells will also be absorbed by the body, making it difficult to maintain stability for a long time
[0009] To sum up, the method of using autologous tissue or cell transplantation to treat soft tissue defects has inherent and insurmountable problems, mainly absorption and trauma in the supply area; however, repairing fat or breast defects with tissue engineering is a current research hotspot. , but breakthrough progress has not yet been made, the main problem is the degradation and absorption of the matrix material due to insufficient blood supply

Method used

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  • Vascularized fat depot based on partition and construction method thereof
  • Vascularized fat depot based on partition and construction method thereof
  • Vascularized fat depot based on partition and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Extraction and Induced Culture of Adipose Stem Cells

[0075] The patient's autologous fat was obtained by liposuction, the adipose tissue was washed with phosphate buffer, and then 0.75g / LI type collagenase was added, shaken and digested at 37°C for 60min, and the digestion was terminated with fetal bovine serum. Centrifuge at 1200r / min for 10min, remove the supernatant and suspended residual tissue, resuspend the cells, add 2 times the volume of red blood cell lysate (NH 4 Cl 154mmol / L+KHCO 3 10mmol / L+EDTA 0.1mmol / L) let stand for 10min, centrifuge and discard the supernatant. Wash with an appropriate amount of PBS for 3 times, and filter through a 200-mesh sieve to obtain adipose tissue stem cells.

[0076] Inoculate the obtained adipose tissue stem cells on 25cm 2 Add 5 mL of high-glucose DMEM medium containing 0.10 volume fraction of fetal bovine serum to the culture bottle, mix well, and place in 37°C, 0.05 volume fraction of CO 2 Cultured in a satur...

Embodiment 2

[0091] Using core-melting technology (patent publication number: 1654028) to prepare two sets of internally connected secondary vascular stents, such as image 3 shown. The first level simulates the biological structure of small blood vessels with an inner diameter of 1000 μm, and the second level structure simulates microvessels with an inner diameter of 300 μm. They are cultivated and trained in a pulsating bioreactor for 2 hours a day for one week, and the circulating fluid is 10 6 pcs / ml smooth muscle cell suspension (the solvent is the corresponding cell culture medium), so that the smooth muscle cells evenly attach to the channel of the vascular stent to form multi-layered smooth muscle cells.

[0092] The fat stem cell microcapsules and growth factor sustained-release microspheres prepared in Example 1 were uniformly mixed with gelatin at room temperature at a volume ratio of 10:1:5, cast into a hemisphere with a diameter of 2 cm, and then cooled to gel; Two sets of gr...

Embodiment 3

[0094] Adipose stem cells cultured routinely were taken using sodium alginate / gelatin system as encapsulation material, and a non-contact high-voltage electrostatic generator (see figure 2 ) to make solid cell microcapsules:

[0095] 1. The sodium alginate solution and the gelatin solution (solvent PBS buffer solution) were mixed to obtain solutions with final concentrations of 1.8% (w / w) and 3% (w / w), respectively.

[0096] 2. Adipose-derived stem cells were treated with 10 6 The density per ml is evenly mixed with the encapsulating solution and then filled into a disposable needle tube with a volume of 10 ml.

[0097] 3. Connect the non-contact high-voltage electrostatic microcapsule generator. The high-voltage electrostatic generating device of this equipment can use SA167-Y (Tianjin) high-voltage electric field generator with an output voltage of 10kV; Syringe pump, using a 5mL syringe, the distance between the needle tip and the copper sheet is 10mm, the advancing spee...

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Abstract

The invention relates to a vascularized adipose tissue based on partition and a construction method thereof. The vascularized adipose tissue of the invention imitates the structure of a natural adipose tissue to coat an adipose cell into a microcapsule and realize partition with a vascular tissue; the construction method thereof comprises: 1) construction of an adipose area; 2) preparation of thecomponents of the blood vessel; 3) construction of the vascularized adipose tissue, which comprises the steps as follows: i) a nutrilit sustained-release microspheres and an adipose tocyst are mixed with a host material and used as a main material, a smooth muscle cell is used as a channel material to construct a 3D structure body provided with a through channel; ii) pulsatile in vitro culture iscarried out on the 3D structure body; and perfusion is carried out with the suspension of an endothelial cell to lead the endothelial cell to be adhered to the inner wall of the channel to form the vascular tissue the outer wall of which is the smooth muscle cell and the inner wall of which is the endothelial cell. Based on the construction method of the invention, the adipose tissue which has adipose and blood vessel distributed by partition and graded vascular structures and can exist in a body for a long period can be obtained.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a partition-based vascularized adipose tissue and a construction method thereof. Background technique [0002] Every year, millions of patients around the world need surgical repair and reconstruction due to soft tissue and breast defects caused by trauma, tumor resection and congenital defects. Breast cancer is one of the most common malignant tumors in women. It is as high as 10% to 15%, and it is also showing a trend of increasing and getting younger year by year. The number of soft tissue and breast defects caused by tumor resection is the largest. According to the American Society of Orthopedic Surgeons, more than 5 million people worldwide underwent breast reconstruction surgery in 2004, 4 million of which were due to tumor removal. . [0003] It is generally believed that an ideal breast filling material should have similar softness, the same osmotic pressure, stable physic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12M3/00B01J13/02A61L27/38C12N5/077
Inventor 姚睿颜永年张人佶王小红林峰栾杰熊卓隋少春张磊卢清萍
Owner TSINGHUA UNIV
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