Cell sheet for tissue repair and bio-artificial tissue engineering, method of producing the same and method of using the same

A cell sheet and cell technology, applied in biochemical equipment and methods, tissue cell/virus culture devices, biochemical instruments, etc., can solve problems such as long degradation time

Active Publication Date: 2012-08-01
THE CHINESE UNIVERSITY OF HONG KONG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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Method used

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  • Cell sheet for tissue repair and bio-artificial tissue engineering, method of producing the same and method of using the same
  • Cell sheet for tissue repair and bio-artificial tissue engineering, method of producing the same and method of using the same
  • Cell sheet for tissue repair and bio-artificial tissue engineering, method of producing the same and method of using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1. Preparation of Cell Sheets

[0093] Rat TDSC stem cells were isolated from rat tendons as described by Rui et al. Tissue Eng Part A 2010; 16(5): 1549-1558. Freeze isolated TDSCs in liquid nitrogen until use. To form cell sheets, TDSC were expanded in growth medium in plastic dishes until confluent. Then, in a humidified cell culture incubator at 10-45 °C, 3-10% CO 2 Under the condition of 1- At 6 weeks, the medium also contained 2-30% fetal bovine serum (FBS), 10-500 U / ml penicillin, 10-500 μg / ml streptomycin and 1-10 mM L-glutamine. The medium containing CTGF and ascorbic acid was replaced every 2-5 days. refer to Figure 7 with Figure 12 , in a humidified incubator at 37 °C, 5% CO 2 Under the condition of , after treating stem cells with ascorbic acid (25μM) and CTGF (25ng / ml) prepared in DMEM containing 2% FBS, the treated cells differentiated, produced abundant extracellular matrix and the resulting cell sheet was elastic. The medium containing ...

Embodiment 2

[0098] Example 2. Formation of bioartificial tendons and ligaments

[0099] refer to Figure 9 , the cell sheets formed with rat TDSC were rolled up and loaded onto U-shaped elastic needles to form bioartificial tendons and ligaments. Referring to FIG. 15, after the cell sheet formed of rat TDSC was transplanted and sutured to the spinal muscle, the cell sheet was further matured. At week 8 after transplantation, cellularity was lower than at week 6, and tendon-like and ligament-like tissues were formed. The transplanted cells were viable, spindle-shaped, aligned longitudinally along the loading axis and embedded between parallel fibrils. At week 8, fibrils were thicker than before transplantation and exhibited collagen birefringence typical of tendons and ligaments.

Embodiment 3

[0100] Example 3. Tissue Repair

[0101] After removal of the patellar-patellar tendon-bone graft in anterior cruciate ligament (ACL) reconstruction, a window-like injury in the patellar tendon can be created by rolling and suturing a cell sheet with active TDSC in the window-like defect. Cell sheets are used to enhance the repair of window-like injuries in the patellar tendon. refer to Figure 16-19 , Suturing cell sheets formed with treated rat TDSC to patellar window defects promoted tendon healing. At week 2, some cells in the defect transplanted with cell sheets became flattened. This phenomenon was not observed in the group without cell sheets. In the group transplanted with cell sheets, more ECMs were observed. The ECM is aligned longitudinally along the tendon and exhibits collagen birefringence typical of ligaments and tendons. At week 2, the transplanted cell sheet was still present in the window-like defect, as shown by ex vivo fluorescence imaging. At week 8,...

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Abstract

Disclosed is a cell sheet for tissue repair and bio-artificial tissue engineering. The cell sheet comprises treated stem cell embedded in its self-secreted extracellular matrix (ECM) and formed a cell sheet. The cell sheet is formed by isolating the stem cell, expanding the stem cell and treating the stem cell with biological factors or factors leading to the production of biological factors, to induce its differentiation, production of extracellular matrix and formation of a cell sheet in vitro. The cell sheet is used as a bioactive material or as an acellular material for the promotion of tissue repairs or used to form a bio-artificial organ for tissue replacement. The cell sheet of the present invention eliminates the need to use scaffolds for cell delivery. The cell sheet facilitates in vivo cell transplantation and provides some tensile mechanical strength for bearing early mechanical load during tissue repair.

Description

field of invention [0001] The present invention relates to a cell sheet, a method for preparing the cell sheet, and a method for using the cell sheet to promote tissue repair and bioartificial tissue engineering. Background of the invention [0002] Musculoskeletal and connective tissue diseases have a devastating impact on society. They are the most common cause of severe chronic long-term pain and loss of physical activity, affecting people of all ages and imposing a substantial cost burden on society. One in three women over the age of 50 suffers a fracture from osteoporosis. Between 23 and 34 million people are injured in road traffic accidents worldwide each year. Back pain is the second leading cause of occupational sick leave. Up to 80% of people suffer from back pain in their lifetime. Osteoarthritis affects more than 135 million people worldwide. Osteoarthritis is the 4th most common cause of health problems in women and the 8th most common cause of health prob...

Claims

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Application Information

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IPC IPC(8): A61L27/38
CPCC12M3/00A61L27/3895A61L27/3834A61P17/00A61P17/02A61P19/00A61P19/02A61P19/08A61P21/00
Inventor 吕宝仪倪明芮云峰
Owner THE CHINESE UNIVERSITY OF HONG KONG
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