Cartilage tissue engineering using cryogenic chondrocytes.
OBJECTIVE
To generate in vitro hyaline cartilage from cryogenically preserved human septal chondrocytes in a simulated microgravity environment on a 3-dimensional biodegradable scaffolding material.


METHODS
In this experiment, cryogenically frozen chondrocytes were thawed and cultured in a monolayer in serum-based chondrocyte media. They were seeded onto 3-dimensional biopolymer scaffolds in a spinner flask. The seeded constructs were then transferred to a bioreactor (an environment of solid-body rotation) for 6 weeks. Chondrocyte growth and extracellular matrix production in the constructs were confirmed by cell count, cell viability, and histologic analysis and by electron microscopy.


RESULTS
Histologic sections stained with hematoxylin-eosin and Alcian blue (for acidic proteoglycans) confirmed the presence of hyaline cartilage in the cartilage constructs. Ultrastructural examination using transmission electron microscopy demonstrated matrix formation and chondrocyte viability.


CONCLUSIONS
This study proves that chondrocytes that are cryogenically stored for extended periods can be used to grow cartilage in vitro. Cryogenically preserved chondrocytes retain their ability to grow in tissue culture, redifferentiate, and produce extracellular matrix.
Gorti
Goutham K
GK
Wound Healing and Tissue Engineering Laboratory, Division of Otolaryngology-Head and Neck Surgery, Stanford University Medical Center, Stanford, Calif., USA.
Lo
Justin
J
Falsafi
Sassan
S
Kosek
Jon
J
Quan
Susan Y
SY
Khuu
Duke T
DT
Koch
R James
RJ
D019149:Bioreactors; D018929:Cell Culture Techniques; D019902:Chondrocytes / Q000166:cytology / Q000502:physiology*; D015925:Cryopreservation; D005109:Extracellular Matrix / Q000502:physiology*; D006801:Humans; D008854:Microscopy, Electron; D009300:Nasal Septum / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D014893:Weightlessness