title: Methodological models for in vitro amplification and maintenance of human articular chondrocytes from elderly patients.
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.
authors: Department of Internal Medicine, University of Florence, Viale Pieraccini 6, 50139 Florence, Italy.
authors: Anna Maria
authors: AM
authors: Carossino
authors: Raffaella
authors: R
authors: Recenti
authors: Roberto
authors: R
authors: Carossino
authors: Elisabetta
authors: E
authors: Piscitelli
authors: Alessia
authors: A
authors: Gozzini
authors: Valentina
authors: V
authors: Martineti
authors: Carmelo
authors: C
authors: Mavilia
authors: Alessandro
authors: A
authors: Franchi
authors: Daniele
authors: D
authors: Danielli
authors: Paolo
authors: P
authors: Aglietti
authors: Antonio
authors: A
authors: Ciardullo
authors: Gianna
authors: G
authors: Galli
authors: Isabella
authors: I
authors: Tognarini
authors: Alberto
authors: A
authors: Moggi Pignone
authors: Mario
authors: M
authors: Cagnoni
authors: Maria Luisa
authors: ML
authors: Brandi
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002358:Cartilage, Articular / Q000166:cytology* / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D004339:Drug Compounding; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D012333:RNA, Messenger / Q000378:metabolism; D012038:Regeneration / Q000502:physiology
chemical_list: D024042:Collagen Type I; D024043:Collagen Type II; D012333:RNA, Messenger