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Alginate hydrogel microcarrier and preparation method thereof

An alginate and hydrogel technology, applied in bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve problems such as adverse effects on karyotype stability of stem cells, effects on cell yield, and dangers of clinical use, etc. Achieve the effects of good cell compatibility, abundant raw material sources, and good mechanical strength

Inactive Publication Date: 2012-10-31
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies have shown that the enzymatic hydrolysis process is likely to adversely affect the stability of stem cell karyotype [(a) Mitalipova MM, Rao RR, Hoyer DM, Johnson JA, Meisner LF, Jones KL, et al. Preserving the genetic integrity of human embryonic stem cells (preserve the genetic integrity of human embryonic stem cells) Nature Biotechnology, 2005, 23: 19-20]
At the same time, when MSCs are used as seed cells for cell therapy or tissue engineering, the incorporation of animal-derived digestive enzyme components may cause potential danger to their clinical use [Zhang J, Skardal A, Prestwich GD. Engineered extracellular matrices with cleavable crosslinkers for cell expansion and easy cell recovery (engineered extracellular matrix that can be used for cell proliferation and cell recovery) Biomaterials, 2008, 29: 4521-4531]
And more importantly, because microcarriers are mostly macroporous structures, a considerable number of cells (sometimes even reaching more than 40%) exist in the internal pores of the carrier after three-dimensional dynamic amplification, so it is difficult to digest them through conventional enzyme digestion methods. Completely digested, so that the final cell yield is affected [Bacel S, Hu WS.Confocal laser scanning microscopy examination of cell distribution in macroporous microcarriers (confocal laser scanning microscope detection of porous microcarrier cell distribution) Biotechnology Progress, 1996, 12: 398-402]

Method used

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  • Alginate hydrogel microcarrier and preparation method thereof
  • Alginate hydrogel microcarrier and preparation method thereof
  • Alginate hydrogel microcarrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Weigh 0.45 g of sodium alginate and add it to 10 mL of phosphate buffer solution with a pH of 5, and stir at room temperature until the sodium alginate is completely dissolved. Take 2g of sodium alginate solution, add 0.6g of EDC, and stir at 4°C for 30min.

[0023] (2) Add 5 g of peanut oil and 0.25 g of Tween 80 aqueous solution with a mass concentration of 25% to the above solution, and stir at room temperature until a uniform suspension is formed. Then 1.5 ml of cystamine aqueous solution with a mass concentration of 37% was added, and the reaction was performed under constant temperature magnetic stirring at 40° C. and 500 rpm for 1 h. The product was suction filtered and washed with absolute ethanol at the same time to obtain alginate hydrogel microcarriers.

[0024] (3) Soak the alginate hydrogel microcarrier that step (2) obtains in the 20ml chitosan solution that the relative molecular mass is 5k, mass concentration is 0.3% for 5h, then in the 50ml phospha...

Embodiment 2

[0028] The raw materials and preparation method are similar to those in Example 1, except that the amount of sodium alginate is changed from 4.5g to 0.6g, and the prepared 0.1g alginate hydrogel microcarrier is 10ml, 20mmol / L at 37°C. The degradation time in cysteine ​​solution is 25-28 hours.

Embodiment 3

[0030] The raw materials and preparation method are similar to Example 2, except that the amount of Tween 80 aqueous solution added is changed from 0.25g to 0.37g, and the prepared 0.1g alginate hydrogel microcarrier is 10ml, 20mmol at 37°C / L cysteine ​​solution degradation time is 25 ~ 28h.

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Abstract

The invention relates to an alginate hydrogel microcarrier and a preparation method thereof. With a molecular structure represented by the formula 1, the alginate hydrogel microcarrier is prepared by the steps of: under the activation of water-soluble carbodiimide, carboxyl in sodium alginate and amino in cystamine undergo an amidation reaction by a suspension crosslinking method so as to generate a chemically crosslinked alginate hydrogel microcarrier, freeze drying the chemically crosslinked alginate hydrogel microcarrier so as to obtain a porous structured microcarrier which is adhered with chitosan, heparin, basic fibroblast growth factors, fibronectins and other components. The alginate hydrogel microcarrier provided in the invention has simple preparation method, abundant raw material source, good degradability in vitro and cellular compatibility. Application of the alginate hydrogel microcarrier of the invention to mesenchymal stem cell amplification in vitro can avoid the damage of a pancreatin digestion process to cells, and can also increase the cell output.

Description

technical field [0001] The invention relates to an alginate hydrogel microcarrier and a preparation method thereof, belonging to high molecular biomedical materials. Background technique [0002] Mesenchymal stem cells (MSCs) show multipotent differentiation potential when cultured in vitro, and are easy to isolate and culture in vitro, and do not express co-stimulatory antigen B7 [Brinchmann JE. Expanding autologous multipotent mesenchymal bone marrow stromal cells (autologous multiple It can expand bone marrow mesenchymal stem cells) Journal of the neurological sciences, 2008, 265: 127-130], so it is an ideal cell source for cell therapy, and has been widely used in the fields of regenerative medicine and tissue engineering. However, the number of MSCs in human tissues is extremely small, accounting for about 0.001% to 0.01% of bone marrow nucleated cells, which is far from meeting the needs of clinical treatment [Croft A, Przyborski S. Mesenchymal stem cells from the bone...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08L5/04C08J9/42C08J9/40C08J9/28C08J7/12C08J3/24C08J3/075C12N5/0775
Inventor 尹玉姬赵爽赵燕燕
Owner TIANJIN UNIV
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