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Copolymerized nano composite hydrogel for intelligent separation of cell sheets as well as preparation method and application thereof

A nanocomposite, cell sheet technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems such as inhibiting cell metabolic activity, slowing down the process of cell detachment, etc., to avoid tissue fibrosis, reaction The effect of simple controllability, good mechanical properties and temperature responsiveness

Inactive Publication Date: 2012-06-27
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Another study reported (Biomaterials, 1995, 16(4): 297–303) that cell desorption is affected by the hydrophobic-hydrophilic transition of PNIPAm and the level of cell metabolism. Lowering the temperature can increase the degree of hydration of PNIPAm, but the temperature is too low It will also inhibit cell metabolism and slow down the process of cell desorption

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] In a sterile laboratory, disperse 0.9 g Laponite XLS in 28.5 g sterile water, stir for 3 h to obtain a uniform and transparent dispersion; then add 14 μL polyethylene glycol macromonomer and continue stirring for 2 h; then add NIPAm monomer 3.4 g and continue to stir for 2 h; finally, deoxygenate the reaction liquid for 5 min, add initiator potassium persulfate (KPS, 20 mg / mL) 1.5 mL and tetramethylethylenediamine (TEMED) 30 μL , stirred evenly, transferred to glass test tubes and reaction molds with a thickness of 2 mm and sealed, and reacted in a biochemical incubator at 20 °C for 24 h to obtain lithium alginate cross-linked copolymer nanocomposite hydrogel. The hydrogel prepared in the glass test tube was used to measure the mechanical properties. The elongation at break of the hydrogel was 1144%, and the breaking strength was 107 kPa. The hydrogel was taken out of the reaction mold, soaked in a large amount of sterile water, swelled and deswelled repeatedly at 20 °C...

Embodiment 2

[0031] This example is the same as Example 1 except for the following features: in a sterile laboratory, disperse 0.9 g of Laponite RDS in 28.5 g of sterile water, stir for 3 h to obtain a uniform and transparent dispersion; then add 43 μL poly Continue to stir the ethylene glycol macromonomer for 2 h; then add 3.4 g of NIPAm monomer and continue to stir for 2 h; finally ventilate and deoxygenate the reaction liquid for 5 min, add the initiator potassium persulfate (KPS, 20 mg / mL) 1.5 mL and 30 μL of tetramethylethylenediamine (TEMED), stirred evenly, transferred to glass test tubes and reaction molds with a thickness of 2 mm and sealed, and reacted in a biochemical incubator at 20 °C for 24 h to obtain lithium algae. linked copolymeric nanocomposite hydrogels. The elongation at break of the hydrogel is 1084%, the breaking strength is 120 kPa, and the phase transition temperature is 35.5 ℃. After 7 days, the number of cells on the surface of the gel increased to 49 × 10 4 in...

Embodiment 3

[0033] This example is the same as Example 1 except for the following features: in a sterile laboratory, disperse 1.8 g of Laponite XLS in 28.4 g of sterile water, stir for 3 h to obtain a uniform and transparent dispersion; then add 72 μL poly Continue to stir the ethylene glycol macromonomer for 2 h; then add 3.4 g of NIPAm monomer and continue to stir for 2 h; finally ventilate and deoxygenate the reaction liquid for 5 min, add the initiator potassium persulfate (KPS, 20 mg / mL) 1.5 mL and 30 μL of tetramethylethylenediamine (TEMED), stirred evenly, transferred to glass test tubes and reaction molds with a thickness of 2 mm and sealed, and reacted in a biochemical incubator at 20 °C for 24 h to obtain lithium algae. linked copolymeric nanocomposite hydrogels. The elongation at break of the hydrogel was 774%, the strength at break was 217 kPa, and the phase transition temperature was 33.2 ℃. After 7 days, the number of cells on the surface of the gel increased to 70 × 10 4 ...

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Abstract

The invention discloses a copolymerized nano composite hydrogel for intelligent separation of cell sheets as well as a preparation method and application of the copolymerized nano composite hydrogel. The preparation method comprises the steps of: dispersing laponite in water, stirring to obtain homogenous and transparent dispersion, then sequentially adding polyethylene glycol macromonomer and N-isopropyl acrylamide monomer, uniformly stirring, removing oxygen, adding an initiator, transferring reaction liquid to a glass test tube and a reaction mold, sealing, and carrying out in-situ radicalpolymerization reaction at 15-25 DEG C to obtain a target product. The prepared copolymerized nano composite hydrogel has good mechanical performance and temperature sensitivity, and can be used for cell culture and rapid desorption of the cell sheets. The copolymerized nano composite hydrogel is suitable for cell growth, and can also realize rapid and automatic desorption of the cell sheets fromthe surface of the hydrogel by reducing the ambient temperature of the gel, thus tissue fibrosis and inflammation generation caused by use of biodegrable supports are avoided, and the damage to extracellular matrix and intercellular connection caused by use of trypsogen is avoided.

Description

technical field [0001] The invention relates to a cell culture carrier in the fields of biomedicine and tissue engineering, in particular to a temperature-sensitive copolymerized nanocomposite hydrogel and its application in cell culture and spontaneous rapid detachment of cell sheets. Background technique [0002] Since the 20th century, poly N - Isopropylacrylamide (PNIPAm) has been widely used in the field of biomedicine and tissue engineering, including controlled release of drugs, gene expression, chromatographic separation of biomolecules, etc., especially, the use of temperature sensitivity of PNIPAm to obtain intact Methods of damaging cell sheets have received widespread attention. Because PNIPAm has both hydrophilic groups and hydrophobic groups in its molecular chain, when the temperature is 32°C higher than its lower critical solution temperature (LCST), the hydrogen bond between the molecular chain and water molecules is destroyed, and the hydrophobic interacti...

Claims

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Application Information

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IPC IPC(8): C08F290/06C08F120/54C08F2/44C08K3/34C08J3/075C08L51/08C12M3/00C12N5/071
Inventor 童真刘丹王涛郑树典刘新星
Owner SOUTH CHINA UNIV OF TECH
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