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  • Mechanical loading of chondrocytes embedded in 3D constructs: in vitro methods for assessment of morphological and metabolic response to compressive strain.

Mechanical loading of chondrocytes embedded in 3D constructs: in vitro methods for assessment of morphological and metabolic response to compressive strain.

Methods in molecular medicine (2004-07-29)
David A Lee, Martin M Knight
ABSTRACT

Mechanical loading of chondrocytes in 3D constructs has been used to investigate mechanotransduction and its potential for stimulating tissue-engineered cartilage repair. This chapter describes the preparation of 3D agarose or alginate constructs seeded with isolated chondrocytes and specific test rigs for applying gross compressive strain to individual constructs on a confocal microscope or for longer term compression of constructs cultured within an incubator. Experimental methods are described to quantify the level of cell deformation and the elaboration of extracellular matrix. The chapter thus provides an introduction to the experimental techniques used to examine chondrocyte mechanotransduction and downstream cell function.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Agarase from Pseudomonas atlantica, lyophilized powder, ≥5,000 units/mg protein (Lowry)
Sigma-Aldrich
Collagenase from Clostridium histolyticum, powder, Suitable for the digestion and isolation of physiologically active pancreatic islet cells, suitable for cell culture
Sigma-Aldrich
Papain from papaya latex, buffered aqueous suspension, 2× Crystallized, ≥16 units/mg protein
Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, reagent grade, ≥98% (TLC)