Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity.
OBJECTIVE
We investigated how aging effects human chondrocyte yield, proliferation, post-expansion chondrogenic capacity, and response to specific growth factors supplemented during expansion.


METHODS
Fifty-two samples of human articular cartilage were harvested from cadavers 20 to 91 years old and grouped into age decades. Cell yields were normalised to tissue wet weight. Cell proliferation rates were calculated during expansion in medium without (CTR) or with TGF beta 1, FGF-2 and PDGF-BB (TFP). Chondrogenic capacity of CTR- and TFP-expanded cells was assessed by cultivation as 3D pellets in a defined serum-free medium, followed by histological, immunohistochemical, biochemical and real-time RT-PCR analyses.


RESULTS
Cell yields were similar in donors up to 40 years of age and significantly lower (1.8-fold) in older donors. Cell proliferation rates in CTR medium significantly decreased after 30 years of age and remained similar in older donors. In the presence of TFP, proliferation rates were higher (up to 3.7-fold) in all age groups and decreased only slightly with age. The glycosaminoglycan (GAG) content of pellets obtained from CTR-expanded cells was not correlated with age. Pellets from TFP-expanded cells reproducibly contained more GAG (up to 3.2-fold) than those from CTR-expanded cells only if donors were younger than 40. Safranin O staining intensity and collagen type II expression and accumulation were consistent with GAG contents.


CONCLUSIONS
Medium supplementation with the growth factor combination TFP during chondrocyte expansion supports higher proliferation rates at any age and higher post-expansion chondrogenic capacity in donors up to 40 years. These findings may be relevant for chondrocyte-based cartilage repair procedures.
Barbero
Andrea
A
Department of Surgery, University Hospital, Basel, Switzerland.
Grogan
Shawn
S
Schäfer
Dirk
D
Heberer
Michael
M
Mainil-Varlet
Pierre
P
Martin
Ivan
I
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000473:pathology* / Q000502:physiology; D000077214:Becaplermin; D002358:Cartilage, Articular / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects; D020219:Chondrogenesis / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D008875:Middle Aged; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis; D023822:Tissue Engineering / Q000379:methods; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D016222:Fibroblast Growth Factor 2; D000077214:Becaplermin