TY - JOUR
T1 - New technique of seeding chondrocytes into microporous poly(L-lactide-co- ε-caprolactone) sponge by cyclic compression force-induced suction
AU - Xie, Jun
AU - Jung, Youngmee
AU - Kim, Soo Hyun
AU - Kim, Young Ha
AU - Matsuda, Takehisa
PY - 2006/7
Y1 - 2006/7
N2 - The initial requirement for a functional engineered cartilage tissue is the effective and reproducible seeding of chondrocytes into the interior of microporous scaffolds. High seeding efficiency, high cell viability, uniform cell distribution, and short operation time are also essential. We devised a new technique of seeding rabbit chondrocytes into microporous poly(L-lactide-co- ε-caprolactone) (PLCL) (porosity, 71-80%; wall thickness, 2 and 6 mm) sponges under compression force-induced suction using a custom-designed loading apparatus. Cell distribution and cell viability were determined using confocal laser scanning microscopy with fluorescent dye-staining techniques. Factors that affect the quality of a cell-seeded construct were studied, namely, the porosity and thickness of sponges and suction cycles. Under 1 cycle of suction, an increase in porosity promoted cell seeding efficiency (CSE; defined as the percentage of the number of cells in the sponges relative to the initial number of cells seeded), cell viability (at 1 day postseeding), and a relatively uniform cell distribution, whereas thick sponges exhibited an inhomogeneous cell distribution irrespective of incubation time. Multiple cycles of suction of 5 and 10 at 0.1 Hz significantly improved the CSE, whereas high cell viability was maintained and even spatial cell distribution was achieved in 1 week. This study revealed that our newly developed cell seeding technique with multiple cycles of suction is a promising approach to inoculating cells into microporous sponges with high CSE, high cell viability, and homogeneous cell distribution.
AB - The initial requirement for a functional engineered cartilage tissue is the effective and reproducible seeding of chondrocytes into the interior of microporous scaffolds. High seeding efficiency, high cell viability, uniform cell distribution, and short operation time are also essential. We devised a new technique of seeding rabbit chondrocytes into microporous poly(L-lactide-co- ε-caprolactone) (PLCL) (porosity, 71-80%; wall thickness, 2 and 6 mm) sponges under compression force-induced suction using a custom-designed loading apparatus. Cell distribution and cell viability were determined using confocal laser scanning microscopy with fluorescent dye-staining techniques. Factors that affect the quality of a cell-seeded construct were studied, namely, the porosity and thickness of sponges and suction cycles. Under 1 cycle of suction, an increase in porosity promoted cell seeding efficiency (CSE; defined as the percentage of the number of cells in the sponges relative to the initial number of cells seeded), cell viability (at 1 day postseeding), and a relatively uniform cell distribution, whereas thick sponges exhibited an inhomogeneous cell distribution irrespective of incubation time. Multiple cycles of suction of 5 and 10 at 0.1 Hz significantly improved the CSE, whereas high cell viability was maintained and even spatial cell distribution was achieved in 1 week. This study revealed that our newly developed cell seeding technique with multiple cycles of suction is a promising approach to inoculating cells into microporous sponges with high CSE, high cell viability, and homogeneous cell distribution.
UR - http://www.scopus.com/inward/record.url?scp=33746687836&partnerID=8YFLogxK
U2 - 10.1089/ten.2006.12.1811
DO - 10.1089/ten.2006.12.1811
M3 - Article
C2 - 16889511
AN - SCOPUS:33746687836
SN - 1076-3279
VL - 12
SP - 1811
EP - 1820
JO - Tissue Engineering
JF - Tissue Engineering
IS - 7
ER -