1.Expression and Regulation of Latent TGF-beta Binding Protein-1 Transcripts and Their Splice Variants in Human Glomerular Endothelial Cells.
Joon Hyeok KWAK ; Ji Su WOO ; Kunyoo SHIN ; Hee Joon KIM ; Hoe Su JEONG ; Dong Cheol HAN ; Sung Il KIM ; Choon Sik PARK
Journal of Korean Medical Science 2005;20(4):628-635
Latent transforming growth factor (TGF)-beta-binding protein (LTBP) is required for the assembly, secretion, matrix association, and activation of latent TGF-beta complex. To elucidate the cell specific expression of the genes of LTBP-1 and their splice variants and the factors that regulate the gene expression, we cultured primary human glomerular endothelial cells (HGEC) under different conditions. Basal expression of LTBP-1 mRNA was suppressed in HGEC compared to WI-38 human embryonic lung fibroblasts. High glucose, H2O2, and TGF-beta1 upregulated and vascular endothelial growth factor (VEGF) further downregulated LTBP-1 mRNA in HGEC. RT-PCR with a primer set for LTBP-1S produced many clones but no clone was gained with a primer set for LTBP-1L. Of 12 clones selected randomly, Sca I mapping and DNA sequencing revealed that only one was LTBP-1S and all the others were LTBP-1S delta 53. TGF-beta1, but not high glucose, H2O2 or VEGF, tended to increase LTBP-1S delta 53 mRNA. In conclusion, HGEC express LTBP-1 mRNA which is suppressed at basal state but upregulated by high glucose, H2O2, and TGF-beta1 and downregulated by VEGF. Major splice variant of LTBP-1 in HGEC was LTBP-1S delta 53. Modification of LTBP-1S delta 53 gene in HGEC may abrogate fibrotic action of TGF-beta1 but this requires confirmation.
*Alternative Splicing
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Amino Acid Sequence
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Cell Line
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Cells, Cultured
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Cloning, Molecular
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Comparative Study
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Endothelial Cells/drug effects/*metabolism
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*Gene Expression Regulation
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Glucose/pharmacology
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Humans
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Hydrogen Peroxide/pharmacology
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Intracellular Signaling Peptides and Proteins/*genetics
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Kidney Glomerulus/cytology
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Protein Isoforms/genetics
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RNA, Messenger/genetics/metabolism
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Research Support, Non-U.S. Gov't
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Reverse Transcriptase Polymerase Chain Reaction
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*Transcription, Genetic
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Transfection
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Transforming Growth Factor beta/pharmacology
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Vascular Endothelial Growth Factor A/pharmacology
2.Guidelines for Manufacturing and Application of Organoids: Kidney
Hyun Mi KANG ; Dong Sung KIM ; Yong Kyun KIM ; Kunyoo SHIN ; Sun-Ju AHN ; Cho-Rok JUNG
International Journal of Stem Cells 2024;17(2):141-146
Recent advancements in organoid technology have led to a vigorous movement towards utilizing it as a substitute for animal experiments. Organoid technology offers versatile applications, particularly in toxicity testing of pharmaceuticals or chemical substances. However, for the practical use in toxicity testing, minimal guidance is required to ensure reliability and relevance. This paper aims to provide minimal guidelines for practical uses of kidney organoids derived from human pluripotent stem cells as a toxicity evaluation model in vitro.