Culture method for composition of periosteal biological scaffold and allogenic seed cells

A technology for biological scaffolds and seed cells, which is applied in the field of bioengineering and can solve problems such as the destruction of extracellular matrix components and structures, the difficulty of cell recombination, and the impact on the biocompatibility of scaffolds.

Inactive Publication Date: 2015-04-15
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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  • Abstract
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Problems solved by technology

[0003] Although the periosteal tissue can effectively remove immunogenic cellular components through the action of physics, chemistry and related enzymes, the processing process will inevitably cause un

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  • Culture method for composition of periosteal biological scaffold and allogenic seed cells
  • Culture method for composition of periosteal biological scaffold and allogenic seed cells
  • Culture method for composition of periosteal biological scaffold and allogenic seed cells

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Embodiment 1

[0026] Example 1 The culture method of rabbit tibia non-immunogenic periosteum bioscaffold and seed cells

[0027] 1. Materials

[0028] Experimental animals: Healthy New Zealand white rabbits, provided by the Animal Experiment Center of Wenzhou Medical University, aged 3 to 4 months, weighing about 2.0 to 2.5 kg, male or female, fed with regular diet and water, and kept in separate cages.

[0029] Main reagents and equipment: Triton-X100 (Triton-X100, Sigma Corporation), sodium dodecyl sulfate (SDS, Sigma Corporation, USA), benzyl xanthoyl fluoride (PMSF, Sigma Corporation, USA), RNase , DNase (US Sigma Company), type Ⅰ collagenase (US GIBCO Company), DNA extraction kit (Dalian TAKARA Company), hydroxyproline (Hyp) test box (Nanjing Kaiji Biotechnology), DAPI staining solution (USA Sigma), CCK-8 (Japanese colleagues), fetal bovine serum (GIBCO, USA), penicillin, streptomycin (Shanghai Boyun Biological Co., Ltd.), DMEM / F-12 medium (GIBCO, USA), constant temperature shaking M...

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Abstract

The invention discloses a culture method for the composition of a periosteal biological scaffold and allogenic seed cells. The culture method comprises the following steps: the non-immunogenic periosteal biological scaffold and the seed periosteal cells are prepared; the non-immunogenic periosteal biological scaffold is sealed after low-temperature drying; after gamma-ray disinfection of the non-immunogenic periosteal biological scaffold, the seed periosteal cells, with the concentration of 1*106 per milliliter, are dripped on the surface of the disinfected scaffold; a culture medium is added after the periosteal cells gradually permeate into the non-immunogenic periosteal biological scaffold together with a suspension, the medium change is carried out every 2 to 3 days, and the in-vitro composite culture lasts for one week. The periosteal biological scaffold obtained through the method has the characteristics of complete removal of immunogenic cells, as well as good preservation of the structure and major components of extracellular matrix; effective composition of the allogenic periosteal cells and the biological scaffold can be achieved, so that a biological composite scaffold material can be provided for tissue engineering researches on bone defects and bone nonunion.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a compound culture method of periosteum biological scaffold and allogeneic seed cells. Background technique [0002] The periosteum has a strong bone-forming ability and plays an important role in fracture healing and repair. The periosteum is also considered to be the initiating factor that promotes the repairing effect of bone graft materials in the grafted area. However, autologous periosteum transplantation has problems such as insufficient donors and damage to the structure and function of the donor site, while allogeneic periosteum transplantation has serious problems such as immune rejection. Therefore, solving the immune rejection problem in allogeneic periosteum will be an important link in realizing periosteum transplantation. The main immunogenic components in tissues are cells, and the removal of cells also means the removal of immunogenicity. The...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36
Inventor 陈雷陈凯林贤丰
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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