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Engineered tissue tracing method and construction method

A construction method and engineering technology, applied in biochemical equipment and methods, tissue culture, recombinant DNA technology, etc.

Inactive Publication Date: 2007-10-24
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the above-mentioned defects existing in the current engineered tissue construction and post-implantation observation methods, the purpose of the present invention is to provide a tracer method for engineered tissues to solve the problem of inoculating different types of cells and identifying different batches of inoculated cells question,

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] To construct tissue engineered cartilage in vitro, a large concentration of chondrocytes or stem cells in the scaffold material [(1~5)×10 7 / ml], in order to produce a cartilage matrix close to normal. At present, when constructing tissue-engineered cartilage in vitro, the cells inoculated into the scaffold material tend to fall off easily, so that the local concentration of the seed cells is not high. Larger increase in seed cell concentration. By applying this technology, different planting batches of seed cells can be marked with different fluorescent proteins, so as to display changes in the number and distribution of cells in the same scaffold material.

[0022] step:

[0023] 1. According to the needs of the engineered tissue cartilage to be constructed, prepare the required seed cells, use chondrocytes or stem cells, and amplify them in large quantities.

[0024] 2. Build by yourself or purchase the required retrovirus as a gene carrier from a biological compa...

Embodiment 2

[0032] Example 2 Tracking the Construction and Regeneration of Engineered Bone and Cartilage Composite Tissue

[0033] The engineered bone and cartilage composite tissue is a composite tissue of engineered cartilage tissue and engineered bone tissue constructed in vitro by engineering histology technology, which is used to repair the defect of the bone and cartilage composite tissue at the joint end. By using this technology, seed cells in different regions can be labeled with different fluorescent proteins, so as to study the migration behavior of different cells and the functional role of composite tissues in vivo.

[0034] step:

[0035] 1. According to the needs of the engineered tissue to be constructed, prepare the required seed cells, use chondrocytes, osteoblasts or stem cells, and amplify them in large quantities.

[0036]2. Build by yourself or purchase the required retrovirus as a gene carrier from a biological company, and then perform a large amount of amplificat...

Embodiment 3

[0045] Example 3 Tracking the construction and regeneration of engineered tissue skin

[0046] In order to cover skin tissue defect wounds well, when constructing engineered skin in vitro, two layers of tissue can be biomimetically constructed, one of which is inoculated with keratinocytes to simulate the function of epidermal tissue, and the other layer is inoculated with fibroblasts to simulate part of the dermis organizational function. This technology can be used to label different fluorescent proteins on seed cells in different regions, so as to study the migration behavior of different cells and the functional role of different seed cells in tissues in vivo.

[0047] step:

[0048] 1. Mass expansion culture of seed cells to be labeled, such as keratinocytes, fibroblasts or stem cells.

[0049] 2. Build by yourself or purchase the required retrovirus as a gene carrier from a biological company, and then perform a large amount of amplification.

[0050] 3. Fluorescent p...

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PUM

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Abstract

The invention discloses a cell tracer method for engineering tissue, which comprises: leading the fluorescent protein in vitro with gene carrier into objective seed cells, obtaining the cell to express fluorescent protein; inoculating the cell into engineering tissue frame, then observing the cell with fluorescence microscope. This method can display clearly cell special their distribution, proliferation and function.

Description

technical field [0001] The invention relates to the technical field of engineered tissue. Background technique [0002] At present, the effective measure for the clinical treatment of serious defects or dysfunction of tissues and organs is tissue and organ transplantation, but this treatment method has many defects, such as: limited source of donor tissues and organs, immune rejection of transplanted tissues and organs, and increased chance of viral infection Wait. [0003] In the 1980s, with the in-depth development of basic disciplines such as cell biology and engineering materials, scientists began to boldly explore new methods to reasonably solve this world medical problem. The concept of "tissue engineering" was born from this, which is to apply the principles and methods of engineering and life sciences to prepare biologically active substitutes to restore, maintain or improve the function of damaged tissues. The use of tissue engineering technology to repair defecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/65C12N5/08C12Q1/04C12N5/10C12N15/85
Inventor 杨柳段小军
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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