The cultured cells were derived from rabbit corneal stroma by explant technique following microdissection and serial passage. The ultrastructural features of fourteenth-passage keratocytes were examined with both scanning and transmission electron microscope. The cells can be divided into activated, intermediate and old cell according to the differences in electron density and surface microvillous pattern. The morphologic characteristics of the cultured keratocytes partially resemble those shown in corneal keratocytes in vivo.
Cells, Cultured ; Corneal Keratocytes ; Corneal Stroma ; Microdissection ; Serial Passage

The cultured cells were derived from rabbit corneal stroma by explant technique following microdissection and serial passage. The ultrastructural features of fourteenth-passage keratocytes were examined with both scanning and transmission electron microscope. The cells can be divided into activated, intermediate and old cell according to the differences in electron density and surface microvillous pattern. The morphologic characteristics of the cultured keratocytes partially resemble those shown in corneal keratocytes in vivo.
Cells, Cultured ; Corneal Keratocytes ; Corneal Stroma ; Microdissection ; Serial Passage

The cultured cells were derived from rabbit corneal stroma by explant technique following microdissection and serial passage. The ultrastructural features of fourteenth-passage keratocytes were examined with both scanning and transmission electron microscope. The cells can be divided into activated, intermediate and old cell according to the differences in electron density and surface microvillous pattern. The morphologic characteristics of the cultured keratocytes partially resemble those shown in corneal keratocytes in vivo.
Cells, Cultured ; Corneal Keratocytes ; Corneal Stroma ; Microdissection ; Serial Passage

Cell cultures of adults bovine retinal pigment epithelium(RPE) were propagated from central and peripheral regions of the same eyes to study the topographical differences in cell growth and to compare the differences in growth rate between two areas. The results obtained were as follows: A regional variation in the morpholgy was observed between the RPE from central and that from peripheral regions. Retinal pigment epithelium from central region attached to culture dish more slowly(average 4 days) than those from peripheral region(average 3.5 days) The growth rate of retinal pigment epithelium declined with serial passage in culture. The growth rate of retinal pigment epithelium from peripheral region at the first generation was highest. And there was a statistical difference in growth rate with passing in generation(P<0.05). This study reveals that growth rate and cell activity of RPE from central region are lower than from peripheral region.
Adult ; Cell Culture Techniques ; Humans ; Macular Degeneration ; Retinal Pigment Epithelium ; Retinaldehyde ; Serial Passage

Cell cultures of adults bovine retinal pigment epithelium(RPE) were propagated from central and peripheral regions of the same eyes to study the topographical differences in cell growth and to compare the differences in growth rate between two areas. The results obtained were as follows: A regional variation in the morpholgy was observed between the RPE from central and that from peripheral regions. Retinal pigment epithelium from central region attached to culture dish more slowly(average 4 days) than those from peripheral region(average 3.5 days) The growth rate of retinal pigment epithelium declined with serial passage in culture. The growth rate of retinal pigment epithelium from peripheral region at the first generation was highest. And there was a statistical difference in growth rate with passing in generation(P<0.05). This study reveals that growth rate and cell activity of RPE from central region are lower than from peripheral region.
Adult ; Cell Culture Techniques ; Humans ; Macular Degeneration ; Retinal Pigment Epithelium ; Retinaldehyde ; Serial Passage

Cell cultures of adults bovine retinal pigment epithelium(RPE) were propagated from central and peripheral regions of the same eyes to study the topographical differences in cell growth and to compare the differences in growth rate between two areas. The results obtained were as follows: A regional variation in the morpholgy was observed between the RPE from central and that from peripheral regions. Retinal pigment epithelium from central region attached to culture dish more slowly(average 4 days) than those from peripheral region(average 3.5 days) The growth rate of retinal pigment epithelium declined with serial passage in culture. The growth rate of retinal pigment epithelium from peripheral region at the first generation was highest. And there was a statistical difference in growth rate with passing in generation(P<0.05). This study reveals that growth rate and cell activity of RPE from central region are lower than from peripheral region.
Adult ; Cell Culture Techniques ; Humans ; Macular Degeneration ; Retinal Pigment Epithelium ; Retinaldehyde ; Serial Passage

Cell cultures of adults bovine retinal pigment epithelium(RPE) were propagated from central and peripheral regions of the same eyes to study the topographical differences in cell growth and to compare the differences in growth rate between two areas. The results obtained were as follows: A regional variation in the morpholgy was observed between the RPE from central and that from peripheral regions. Retinal pigment epithelium from central region attached to culture dish more slowly(average 4 days) than those from peripheral region(average 3.5 days) The growth rate of retinal pigment epithelium declined with serial passage in culture. The growth rate of retinal pigment epithelium from peripheral region at the first generation was highest. And there was a statistical difference in growth rate with passing in generation(P<0.05). This study reveals that growth rate and cell activity of RPE from central region are lower than from peripheral region.
Adult ; Cell Culture Techniques ; Humans ; Macular Degeneration ; Retinal Pigment Epithelium ; Retinaldehyde ; Serial Passage

Due to the increased frequency of interspecies transmission of avian influenza viruses, studies designed to identify the molecular determinants that could lead to an expansion of the host range have been increased. A variety of mouse-based mammalian-adaptation studies of avian influenza viruses have provided insight into the genetic alterations of various avian influenza subtypes that may contribute to the generation of a pandemic virus. To date, the studies have focused on avian influenza subtypes H5, H6, H7, H9, and H10 which have recently caused human infection. Although mice cannot fully reflect the course of human infection with avian influenza, these mouse studies can be a useful method for investigating potential mammalian adaptive markers against newly emerging avian influenza viruses. In addition, due to the lack of appropriate vaccines against the diverse emerging influenza viruses, the generation of mouse-adapted lethal variants could contribute to the development of effective vaccines or therapeutic agents. Within this review, we will summarize studies that have demonstrated adaptations of avian influenza viruses that result in an altered pathogenicity in mice which may suggest the potential application of mouse-lethal strains in the development of influenza vaccines and/or therapeutics in preclinical studies.
Animals ; Host Specificity ; Humans ; Influenza A virus ; Influenza in Birds* ; Influenza Vaccines ; Methods ; Mice ; Orthomyxoviridae ; Pandemics ; Serial Passage ; Vaccination ; Vaccines* ; Virulence

We have reported RPS-Vax system by introducing multiple cloning site (MCS) and 3C-protease cutting site at the N-terminal end of the poliovirus Sabin 1 cDNA. Potential vaccine genes can be easily introduced into recombinant polioviral genome and expressed during the viral replication as a part of virus polyprotein and subsequently processed from the mature viral protein by the poliovirus-specific 3C-protease. However, these poliovirus vector-mediated chimeric viral vaccine was not efficient to induce the cell-mediated immunity because of its rapid cytolytic capacity. In order to make CTL-inducing vaccine vector, we integrated a protein transduction domain (PTD) into the pRPS-Vax vector system right ahead of the MCS, named RPS-Vax/PTD. We have incorporated the HCV core (N-terminal 100aa) antigen into the MCS of pRPSvax-PTD vector, followed by production of chimeric virus, named RPSvax-PTD/HCVc. The chimeric virus was genetically stable during the serial passages. Replication capacity of the RPSvax-PTD/HCVc was 1~2 log lower than that of RPS-Vax control virus. These chimeric virus was very efficient to inducing antigen-specific IgG2a in the immunized mice, implying that the recombinant virus has a capacity to induce HCV-specific Th1 type immunity in the immunized animals or humans.
Animals ; Clone Cells ; Cloning, Organism ; DNA, Complementary ; Genome ; Hepatitis C* ; Hepatitis* ; Humans ; Immunity, Cellular ; Immunoglobulin G ; Mice ; Poliovirus ; Serial Passage

Poliovirus Sabin 1 strain has its own special features that make it a particularly attractive live recombinant mucosal vaccine vehicle. Sabin 1 cDNA was manipulated to have multiple cloning site and viral specific 3C-protease cutting site at the N-terminal end of the polyprotein, named RPS-vax system HIV-1 V3- and principal neutralizing domain (PND)-concatamers were successfully cloned into the multiple cloning site of the vector system and produced expected chimeric viruses by transfection of their RNA transcripts into HeLa cells. These chimeric viruses have shown to express introduced HIV-1 subgenome concatamers efficiently during their replication in the infected HeLa cells. Expressed proteins were confirmed to retain the wild type structures at least in parts. Replication capacity of the chimeric viruses was slightly lower than that of wild type Sabin 1 likely to be due to delay in processing steps during their replication. Differing from the virulent Mahoney vectors, the rec-Sabin 1 chimeric viruses maintained the foreign gene stably during the serial passages. These chimeric viruses have also shown to be able to induce specific humoral immunity to the introduced vaccine proteins when inoculated into the poliovirus receptor-expressing transgenic (Tg-PVR) mice. Antiserum obtained from the immunized transgenic mice showed to have neutralizing capacity to HIV-1 in vitro. These results strongly suggest that the chimeric viruses expressing HIV-1 vaccine epitopes can be used as a good live mucosal vaccine candidate against AIDS.
Animals ; Clone Cells ; Cloning, Organism ; DNA, Complementary ; Epitopes ; HeLa Cells ; HIV* ; HIV-1* ; Humans* ; Immunity, Humoral ; Mice ; Mice, Transgenic ; Poliovirus* ; RNA ; Serial Passage ; Transfection