Skip to Content
Merck
  • A Streamlined Method for the Preparation of Growth Factor-enriched Thermosensitive Hydrogels from Soft Tissue.

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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Zinc sulfate heptahydrate, BioReagent, suitable for cell culture
Sigma-Aldrich
Oil Red O, certified by the Biological Stain Commission
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Guanidine hydrochloride, ≥98%
Sigma-Aldrich
Benzamidine hydrochloride hydrate, ≥99%
Sigma-Aldrich
N-Ethylmaleimide, crystalline, ≥98% (HPLC)
Roche
DNase I, from bovine pancreas