Methodological models for in vitro amplification and maintenance of human articular chondrocytes from elderly patients |
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Authors: | Anna Maria Carossino Raffaella Recenti Roberto Carossino Elisabetta Piscitelli Alessia Gozzini Valentina Martineti Carmelo Mavilia Alessandro Franchi Daniele Danielli Paolo Aglietti Antonio Ciardullo Gianna Galli Isabella Tognarini Alberto Moggi Pignone Mario Cagnoni Maria Luisa Brandi |
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Institution: | (1) Departments of Internal Medicine, University of Florence, Viale Pieraccini, 6, 50139 Florence, Italy;(2) Departments of Human Pathology and Oncology, University of Florence, Florence, Italy;(3) Departments of Orthopedics, University of Florence, Florence, Italy;(4) DeGene Spin-off, University of Florence, Florence, Italy |
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Abstract: | Articular cartilage defects, an exceedingly common problem closely correlated with advancing age, is characterized by lack
of spontaneous resolution because of the limited regenerative capacity of adult articular chondrocytes. Medical and surgical
therapies yield unsatisfactory short-lasting results. Recently, cultured autologous chondrocytes have been proposed as a source
to promote repair of deep cartilage defects. Despite encouraging preliminary results, this approach is not yet routinely applicable
in clinical practice, but for young patients. One critical points is the isolation and ex vivo expansion of large enough number
of differentiated articular chondrocytes. In general, human articular chondrocytes grown in monolayer cultures tend to undergo
dedifferentiation. This reversible process produces morphological changes by which cells acquire fibroblast-like features,
loosing typical functional characteristics, such as the ability to synthesize type II collagen. The aim of this study was
to isolate human articular chondrocytes from elderly patients and to carefully characterize their morphological, proliferative,
and differentiative features. Cells were morphologically analyzed by optic and transmission electron microscopy (TEM). Production
of periodic acid-schiff (PAS)-positive cellular products and of type II collagen mRNA was monitored at different cellular
passages. Typical chondrocytic characteristics were also studied in a suspension culture system with cells encapsulated in
alginate-polylysine-alginate (APA) membranes. Results showed that human articular chondrocytes can be expanded in monolayers
for several passages, and then microencapsulated, retaining their morphological and functional characteristics. The results
obtained could contribute to optimize expansion and redifferentiation sequences for applying cartilage tissue engineering
in the elderly patients. |
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Keywords: | Human articular chondrocytes Type I collagen Type II collagen Alginate Cell therapy |
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