Alginate hydrogel microcarrier and preparation method thereof

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...

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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]

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