Methodological models for in vitro amplification and maintenance of human articular chondrocytes from elderly patients.
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.
Carossino
Anna Maria
AM
Department of Internal Medicine, University of Florence, Viale Pieraccini 6, 50139 Florence, Italy.
Recenti
Raffaella
R
Carossino
Roberto
R
Piscitelli
Elisabetta
E
Gozzini
Alessia
A
Martineti
Valentina
V
Mavilia
Carmelo
C
Franchi
Alessandro
A
Danielli
Daniele
D
Aglietti
Paolo
P
Ciardullo
Antonio
A
Galli
Gianna
G
Tognarini
Isabella
I
Moggi Pignone
Alberto
A
Cagnoni
Mario
M
Brandi
Maria Luisa
ML
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
D024042:Collagen Type I; D024043:Collagen Type II; D012333:RNA, Messenger