A high-efficiency somatic embryogenesis protocol of Japanese larch （Larix leptolepis Gordon） has been established in our investigation. Calli were induced from immature zygotic embryos of female cones ofL. leptolepi...A high-efficiency somatic embryogenesis protocol of Japanese larch （Larix leptolepis Gordon） has been established in our investigation. Calli were induced from immature zygotic embryos of female cones ofL. leptolepis and then subcultured regularly on to a modified Gupta and Durzan （DCR） basal medium for 5 years. Embryogenic tissues showed distinct morphological changes dur- ing somatic embryo development when they were transferred to a maturation medium supplemented with abscisic acid （ABA） com- pared with the morphology in a medium lacking ABA. Histological observations indicated that polyembryony was a characteristic feature during early embryogeny and somatic embryos at later stages showed normal histodifferentiation. In addition, histochemical analysis revealed that abundant starch granules and proteins accumulated in mature embryos, indicating that they played important roles in the development and regeneration of normal plantlets from somatic embryos on hormone-free germination media展开更多
Background Efficient cell adhesion and proliferation is a central issue in cell-based tissue engineering, which offers great promise for repair of urethral defects or strictures. This study evaluated the adhesion and ...Background Efficient cell adhesion and proliferation is a central issue in cell-based tissue engineering, which offers great promise for repair of urethral defects or strictures. This study evaluated the adhesion and growth of rabbit uroepithelium on a surface-modified three-dimensional poly-L-lactic acid （PLLA） scaffold.Methods Urethral mucosa were harvested from male New Zealand rabbits and the urothelium were dissociated and then cultured. Immunocytochemistry on cultured uroepithelium for pancytokeratin and uroplakin Ⅱ and TE-7 confirmed pure populations. After in vitro proliferation, cells were seeded onto a surface-modified urethral scaffold with non-knitted filaments. The morphology and viability of the cells were examined by immunohistochemical and fluorescence staining.Inverted and scanning microscopes were used to document cell growth and adhesion.Results Three to five days after primary culture, the uroepithelial cells gradually became confluent, assuming a cobblestone pattern. The filaments of the urethral scaffold had excellent biocompatibility and allowed growth of the uroepithelium, without affecting viability. The uroepithelial cells adhered to and grew well on the scaffold. After 3-7 days,the cells grew vigorously and meshes of the scaffold were full of uroepitheliums.Conclusions The surface-modified urethral scaffold with non-knitted filaments allows the growth of uroepithelium and can serve as a carrier for the tissue engineering of urethra.展开更多
基金the National Natural Science Foundation of China （Grant No. 30471366） the National Science Fund of China for Distinguished Young Scholars （30225005）.
文摘A high-efficiency somatic embryogenesis protocol of Japanese larch （Larix leptolepis Gordon） has been established in our investigation. Calli were induced from immature zygotic embryos of female cones ofL. leptolepis and then subcultured regularly on to a modified Gupta and Durzan （DCR） basal medium for 5 years. Embryogenic tissues showed distinct morphological changes dur- ing somatic embryo development when they were transferred to a maturation medium supplemented with abscisic acid （ABA） com- pared with the morphology in a medium lacking ABA. Histological observations indicated that polyembryony was a characteristic feature during early embryogeny and somatic embryos at later stages showed normal histodifferentiation. In addition, histochemical analysis revealed that abundant starch granules and proteins accumulated in mature embryos, indicating that they played important roles in the development and regeneration of normal plantlets from somatic embryos on hormone-free germination media
基金This work was supported by the grants from the National Natural Science Foundation of China （No. 81070555） and the Major Program of Clinical High and New Technology Research of Hospital.
文摘Background Efficient cell adhesion and proliferation is a central issue in cell-based tissue engineering, which offers great promise for repair of urethral defects or strictures. This study evaluated the adhesion and growth of rabbit uroepithelium on a surface-modified three-dimensional poly-L-lactic acid （PLLA） scaffold.Methods Urethral mucosa were harvested from male New Zealand rabbits and the urothelium were dissociated and then cultured. Immunocytochemistry on cultured uroepithelium for pancytokeratin and uroplakin Ⅱ and TE-7 confirmed pure populations. After in vitro proliferation, cells were seeded onto a surface-modified urethral scaffold with non-knitted filaments. The morphology and viability of the cells were examined by immunohistochemical and fluorescence staining.Inverted and scanning microscopes were used to document cell growth and adhesion.Results Three to five days after primary culture, the uroepithelial cells gradually became confluent, assuming a cobblestone pattern. The filaments of the urethral scaffold had excellent biocompatibility and allowed growth of the uroepithelium, without affecting viability. The uroepithelial cells adhered to and grew well on the scaffold. After 3-7 days,the cells grew vigorously and meshes of the scaffold were full of uroepitheliums.Conclusions The surface-modified urethral scaffold with non-knitted filaments allows the growth of uroepithelium and can serve as a carrier for the tissue engineering of urethra.
