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Injection type vascularized adipose tissue and construction method thereof

A technology of adipose tissue and vascularization, applied in the field of bioengineering, can solve the problems of graft degradation, absorption or necrosis, difficulty in stabilization, hard breast tissue of synthetic polymer materials, etc.

Active Publication Date: 2011-03-30
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the main problem is: on the one hand, the commonly used synthetic polymer materials are too hard compared to breast tissue; on the other hand, if the blood supply cannot be established in time, the degradable polymer materials of composite cells will also be absorbed by the body, making it difficult to maintain stability for a long time
Adipose tissue engineering combined with autologous stem cell technology provides a promising direction, but breakthroughs have not yet been made. The biggest difficulty lies in graft degradation, resorption or necrosis caused by insufficient blood supply

Method used

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  • Injection type vascularized adipose tissue and construction method thereof
  • Injection type vascularized adipose tissue and construction method thereof
  • Injection type vascularized adipose tissue and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The preparation of embodiment 1 fat cell microsphere

[0063] The patient's autologous fat was obtained by liposuction, the adipose tissue was washed with phosphate buffer, and then 0.75g / L type I 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.

[0064] 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 saturated humi...

Embodiment 2

[0071] Example 2 Construction of In Vitro Vascular Structure

[0072] Induction of adipose-derived stem cells into smooth muscle cells: Take adipose-derived stem cells routinely cultured between the fourth and tenth passages, and use smooth muscle-forming medium (M-199 medium containing 5 ng / mL TGF-β1 and 50 ng / mL PDGF-BB) CO at 37°C with a volume fraction of 0.05 2 Cultured in a saturated humidity incubator for 10 days. collect 10 8 smooth muscle cells were centrifuged and pelleted for later use.

[0073] The collagen stock solution was diluted to slightly more than 1 mg / ml with smooth muscle culture medium, then neutralized with 1 mM NaOH and finally adjusted to 1 mg / ml. will be 10 8 A smooth muscle cell was added to the collagen solution and mixed uniformly as the coating material.

[0074] Mix 1ml of the coating material with 1ml of fat cell microspheres evenly and put them into a centrifuge tube. At this time, the cell microspheres sink to the bottom due to their hig...

Embodiment 3

[0078] Angiogenesis growth factor (bFGF and VEGF) sustained-release microspheres were prepared by a non-contact high-voltage electrostatic generator.

[0079] 1. Sodium alginate is prepared into a 1.5% (w / w) solution with PBS buffer solution.

[0080] 2. Mix bFGF and VEGF at a concentration of 2ng / ml and 10ng / ml with the encapsulation solution evenly, and then fill it into a disposable needle tube with a volume of 10ml.

[0081] 3. Connect the non-contact high-voltage electrostatic microcapsule generator, the working voltage is 7kV, the distance between the needle tip and the copper sheet is 20mm, the advancing speed is 30ml / h, the inner diameter of the needle is 191μm, and the collector is a disposable plastic culture with a diameter of 60mm Dish, the solidification solution is 100mmol / L barium chloride solution.

[0082] 4. The growth factor microspheres were collected within 5 minutes, washed twice with normal saline, and then observed and recorded. The shape of the cytoki...

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Abstract

The invention relates to a method for constructing a blood vessel, in particular to an injection type vascularized adipose tissue and a construction method thereof. In the invention, autologous cells are used for simulating a natural vascular tissue, and a natural biological material is used for packaging tissue cells to form microballoons; smooth muscle cells and endothelial cells are compoundedto form a blood vessel structure by utilizing gaps of the microballoons as blood vessel forming channels to obtain an artificial tissue with a blood vessel structure; by simulating the natural vascular tissue, adipose stem cells are packaged into the natural material to prepare the microballoons; and a vascularized factor slow release system is combined to construct the injection type vascularized adipose tissue. According to the invention, the injection type vascularized adipose tissue can be constructed in vitro or in vivo and used for repairing small and large soft tissues, can stably exist in the human body for a long time, has small operational injury and quick postoperative recovery and also can be used for an in vitro biochemically or pharmacally researched model. The method for constructing the blood vessel and the artificial tissue also can be used for other artificial tissues, such as liver tissues, cartilaginous tissues, and the like.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to an injection-type vascularized adipose tissue based on autologous stem cells and a construction method thereof. Background technique [0002] Every year, millions of patients worldwide require surgical repair and reconstruction of soft tissue, facial and breast defects caused by trauma, tumor resection and congenital defects. Because breast cancer is one of the most common malignant tumors in women, the incidence rate 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 fat soft tissue filling material should ha...

Claims

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Application Information

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IPC IPC(8): C12N5/08A61L27/38A61F2/06B01J13/02
Inventor 姚睿颜永年张人佶栾杰王小红林峰熊卓张磊卢清萍
Owner TSINGHUA UNIV
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