Combination of baculovirus-mediated gene transfer and rotating-shaft bioreactor for cartilage tissue engineering.
We have previously demonstrated efficient baculovirus transduction of rat chondrocytes in 6-well plates. To further explore the potential of baculovirus in cartilage tissue engineering, the baculovirus-transduced chondrocytes were seeded into porous scaffolds and cultivated in a rotating-shaft bioreactor (RSB) which was developed for two-phase cultivation of tissue engineered cartilage. The baculovirus transduction resulted in efficiencies up to 90%, and affected neither cell adhesion to the scaffolds nor cell survival in the RSB. After 4-week RSB cultivation, the transduced cells remained highly differentiated and grew into constructs that resembled the untransduced constructs with regard to gross appearance, construct size, cell morphology, cell spatial distribution, glycosaminoglycan and collagen production and deposition. Importantly, baculovirus transduction did not alter the expression of chondrocytic genes. These data confirmed that baculovirus transduction neither harms chondrocytes nor retards the formation of cartilage-like tissues in the RSB, thus implicating the potentials of combining baculovirus-mediated gene transfer with RSB cultivation in in vitro cartilage tissue engineering.
Chen
Huang-Chi
HC
Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan.
Lee
Hsiao-Ping
HP
Ho
Yi-Chen
YC
Sung
Ming-Lun
ML
Hu
Yu-Chen
YC
D053674:Aggrecans; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D019149:Bioreactors; D002356:Cartilage / Q000166:cytology*; D002452:Cell Count; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis / Q000235:genetics; D003094:Collagen / Q000032:analysis / Q000235:genetics; D024043:Collagen Type II / Q000235:genetics; D005109:Extracellular Matrix / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression / Q000235:genetics; D017781:Genes, Immediate-Early / Q000235:genetics; D006016:Glycolates / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019344:Lactic Acid; D037181:Lectins, C-Type / Q000032:analysis / Q000235:genetics; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018411:Spodoptera; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection / Q000379:methods*
C508321:Acan protein, rat; D053674:Aggrecans; D011508:Chondroitin Sulfate Proteoglycans; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D006016:Glycolates; D037181:Lectins, C-Type; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D003094:Collagen