Adult human articular chondrocytes in a microcarrier‐based culture system: expansion and redifferentiation

Abstract

Expanding human chondrocytes in vitro while maintaining their ability to form cartilage remains a key challenge in cartilage tissue engineering. One promising approach to address this is to use microcarriers as substrates for chondrocyte expansion. While microcarriers have shown beneficial effects for expansion of animal and ectopic human chondrocytes, their utility has not been determined for freshly isolated adult human articular chondrocytes. Thus, we investigated the proliferation and subsequent chondrogenic differentiation of these clinically relevant cells on porous gelatin microcarriers and compared them to those expanded using traditional monolayers. Chondrocytes attached to microcarriers within 2 days and remained viable over 4 weeks of culture in spinner flasks. Cells on microcarriers exhibited a spread morphology and initially proliferated faster than cells in monolayer culture, however, with prolonged expansion they were less proliferative. Cells expanded for 1 month and enzymatically released from microcarriers formed cartilaginous tissue in micromass pellet cultures, which was similar to tissue formed by monolayer-expanded cells. Cells left attached to microcarriers did not exhibit chondrogenic capacity. Culture conditions, such as microcarrier material, oxygen tension, and mechanical stimulation require further investigation to facilitate the efficient expansion of clinically relevant human articular chondrocytes that maintain chondrogenic potential for cartilage regeneration applications. ß

Cell expansion of human articular chondrocytes on macroporous gelatine scaffolds—impact of microcarrier selection on cell proliferation

“…The ability of this class of scaffold to support human chondrocyte growth is well known, yet to our knowledge this is the first time that a comparison between these four specific microcarriers has been conducted (Huss et al, 2007;Malda et al, 2003a;Pettersson et al, 2009;Schrobback et al, 2011).…”

“…We have previously proposed the use of gelatine microcarriers in combination with platelet rich plasma as a delivery system for tissue engineering of cartilage using both human dermal fibroblasts, subjected to chondrogenic induction media, and human articular chondrocytes (Sommar et al, 2009;. Other investigators have also used this type of microcarrier in combination with human chondrocytes (Malda et al, 2003a;Pettersson et al, 2009;Huss et al, 2007;Schrobback et al, 2011).…”

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

“…10 Chondrocytes derived from natural sources or from MSCs differentiating toward chondrocytes are commonly used as a cell source in cartilage tissue engineering; however, there are unmet challenges in expanding and maintaining chondrocytes (e.g., the composition of the construct and the regulation of culture conditions need careful monitoring). 13,14,50,56,57 In addition, chondrogenic growth factors play a key role in chondrocyte differentiation and cartilage generation. 58 The most challenging aspect of cartilage tissue engineering is to replicate the natural compressive behavior of cartilage under mechanical loading conditions and its porous structure for cell nutrients.…”

Induction of Re-Differentiation of Passaged Rat Chondrocytes Using a Naturally Obtained Extracellular Matrix Microenvironment

“…A spinner-flask culture is useful in the induction of re-differentiation of dedifferentiated chondrocytes, or in the expansion of microcarrier-attached chondrocytes. 11,12 Furthermore, a hypoxic culture condition with a low oxygen content is also believed to be a factor in promoting chondrocyte re-differentiation. 13 In this study, we obtained cell-derived matrices (CDMs) from in vitro-cultured cells-fibroblasts, preosteoblasts, and chondrocytes-and utilized them as a culture template to evaluate the effect on the induction of re-differentiation of dedifferentiated chondrocytes in a 2D monolayer culture and 3D pellet culture.…”