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Merck

A Streamlined Method for the Preparation of Growth Factor-enriched Thermosensitive Hydrogels from Soft Tissue.

Bio-protocol (2017-02-05)
Christopher J Poon, Shaun S Tan, Sholeh W Boodhun, Keren M Abberton, Wayne A Morrison
RESUMEN

Hydrogels are an ideal medium for the expansion of cells in three dimensions. The ability to induce cell expansion and differentiation in a controlled manner is a key goal in tissue engineering. Here we describe a detailed method for producing hydrogels from soft tissues with an emphasis on adipose tissue. In this method, soluble, extractable proteins are recovered from the tissue and stored while the remaining insoluble tissue is processed and solubilised. Once the tissue has been sufficiently solubilised, the extracted proteins are added. The resulting product is a thermosensitive hydrogel with proteins representative of the native tissue. This method addresses common issues encountered when working with some biomaterials, such as high lipid content, DNA contamination, and finding an appropriate sterilisation method. Although the focus of this article is on adipose tissue, using this method we have produced hydrogels from other soft tissues including muscle, liver, and cardiac tissue.

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Medio de Eagle modificado de Dulbecco, glucosa elevada, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
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DNasa I, from bovine pancreas
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Guanidina hydrochloride, ≥98%
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Sulfato de cinc heptahidratado, BioReagent, suitable for cell culture
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N-Ethylmaleimide, crystalline, ≥98% (HPLC)
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Oil Red O, certified by the Biological Stain Commission
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