Purpose:To study preliminarily induced differentiation of embryonic stem cells intocorneal epithelial cells in vitro.Methods: Murine embryonic stem cells were co-cultured with Rabbit limbal cornealepithelial cells in ...Purpose:To study preliminarily induced differentiation of embryonic stem cells intocorneal epithelial cells in vitro.Methods: Murine embryonic stem cells were co-cultured with Rabbit limbal cornealepithelial cells in Transwell system to induce differentiation. Mophological andimmunohistochemical examination were implemented.Results: The induced cells from embryonic stem cells have an epithelial appearance.The cells formed a network and were confluent into film gradually after beingco-cultured with rabbit limbal corneal epithelial cells for 24 ～ 96 hours. The cells rangedmosaic structure and localized together with clear rim. Most of the cells showedpolygonal appearance. Transmission electron microscope showed lots of microvilli on thesurface of induced cells and tight junctions between them. These epithelial-like cellsexpressed the corneal epithelial cell specific marker cytokeratin3/cytokeratinl2.Conclusion: The potential mechanism of the differentiation of murine embryonic stemcells into corneal epithelial cells induced by limbal corneal epithelial cell-derivedinducing activity is to be further verified.展开更多
Reverse transcription-polymerase chain reaction (RT-PCR) was performed to isolate a cDNA clone using specific primers designed based on the barley (Hordeum vulgare L.) Mlo gene cDNA sequence.A full-length cDNA encodin...Reverse transcription-polymerase chain reaction (RT-PCR) was performed to isolate a cDNA clone using specific primers designed based on the barley (Hordeum vulgare L.) Mlo gene cDNA sequence.A full-length cDNA encoding an Mlo-like protein was isolated and characterized in a Triticum aestivum L.-Haynaldia villosa L. 6VS/6AL translocation line. The putative protein consists of 534 amino acid residues,which contain a nuclear localization motif (NLS), nine casein kinase Ⅱ motifs (S/T-X-X-D/E) and seven protein kinase C motifs (S/T-X-R/K). It is highly homologous to other plant Mlo proteins. Thus, this clone was designated as Ta-Mlo (GenBank accession No. AF384144). Northern blotting analysis showed that the transcription of Ta-Mlo was enhanced slightly by Blumeria graminis (DC) EO Speer f. sp. tritici. Western blotting analysis showed that the protein expression product of the Ta-Mlo gene in wheat seedling leaves is a membrane-bound protein. The protein could be induced by B. graminis. Southern blotting analysis indicated that there is one copy of the Ta-Mlo gene in each wheat genome. Ta-Mlo was localized on specific chromosomal regions of 2AL, 2BL, and 2DL in wheat.展开更多
基金This research was performed with a support from the National Basic Science Research and Development Grants(973)(G1999054301)Natural Sciences Foun- dation of China(No.39770789)Natural Sciences Foundeation of Guangdong Province (No.990093).
文摘Purpose:To study preliminarily induced differentiation of embryonic stem cells intocorneal epithelial cells in vitro.Methods: Murine embryonic stem cells were co-cultured with Rabbit limbal cornealepithelial cells in Transwell system to induce differentiation. Mophological andimmunohistochemical examination were implemented.Results: The induced cells from embryonic stem cells have an epithelial appearance.The cells formed a network and were confluent into film gradually after beingco-cultured with rabbit limbal corneal epithelial cells for 24 ～ 96 hours. The cells rangedmosaic structure and localized together with clear rim. Most of the cells showedpolygonal appearance. Transmission electron microscope showed lots of microvilli on thesurface of induced cells and tight junctions between them. These epithelial-like cellsexpressed the corneal epithelial cell specific marker cytokeratin3/cytokeratinl2.Conclusion: The potential mechanism of the differentiation of murine embryonic stemcells into corneal epithelial cells induced by limbal corneal epithelial cell-derivedinducing activity is to be further verified.
文摘Reverse transcription-polymerase chain reaction (RT-PCR) was performed to isolate a cDNA clone using specific primers designed based on the barley (Hordeum vulgare L.) Mlo gene cDNA sequence.A full-length cDNA encoding an Mlo-like protein was isolated and characterized in a Triticum aestivum L.-Haynaldia villosa L. 6VS/6AL translocation line. The putative protein consists of 534 amino acid residues,which contain a nuclear localization motif (NLS), nine casein kinase Ⅱ motifs (S/T-X-X-D/E) and seven protein kinase C motifs (S/T-X-R/K). It is highly homologous to other plant Mlo proteins. Thus, this clone was designated as Ta-Mlo (GenBank accession No. AF384144). Northern blotting analysis showed that the transcription of Ta-Mlo was enhanced slightly by Blumeria graminis (DC) EO Speer f. sp. tritici. Western blotting analysis showed that the protein expression product of the Ta-Mlo gene in wheat seedling leaves is a membrane-bound protein. The protein could be induced by B. graminis. Southern blotting analysis indicated that there is one copy of the Ta-Mlo gene in each wheat genome. Ta-Mlo was localized on specific chromosomal regions of 2AL, 2BL, and 2DL in wheat.
