1255f12f6a7cc7196de2ca7747ed6aec pmc_10295744.pdf b4426f47aa7096daa9d043192c456e10101441d9 pmc_10295744.pdf e3e43aa54d9b65eaec9b1fa7fc279d4ccba275237cc2e6cc749c54efe989f617 pmc_10295744.pdf Title: Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues Subject: Tritiated thymidine autoradiography, 5-bromo-2'-deoxyuridine (BrdU) 5-chloro-2'-deoxyuridine (CldU), 5-iodo-2'-deoxyuridine (IdU), and 5-ethynyl-2'-deoxyiridine (EdU) labeling have been used for identifying the fraction of cells undergoing the S-phase of the cell cycle and to follow the fate of these cells during the embryonic, perinatal, and adult life in several species of vertebrate. In this current review, I will discuss the dosage and times of exposition to the aforementioned thymidine analogues to label most of the cells undergoing the S-phase of the cell cycle. I will also show how to infer, in an asynchronous cell population, the duration of the G1, S, and G2 phases, as well as the growth fraction and the span of the whole cell cycle on the base of some labeling schemes involving a single administration, continuous nucleotide analogue delivery, and double labeling with two thymidine analogues. In this context, the choice of the optimal dose of BrdU, CldU, IdU, and EdU to label S-phase cells is a pivotal aspect to produce neither cytotoxic effects nor alter cell cycle progression. I hope that the information presented in this review can be of use as a reference for researchers involved in the genesis of tissues and organs. Keywords: thymidine analogues; S-phase; cell cycle parameters; cell kinetics; toxicity Author: Joaquín Martí-Clúa Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Jun 20 06:16:40 2023 CEST ModDate: Tue Jun 20 06:22:27 2023 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1605894 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YXOWQY+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 GGCXGL+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 OIJAQU+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 MVPHJQ+CMSY10 Type 1 Builtin yes yes no 23 0 LPYGPS+VnURWPalladioL-Bold Type 1 Custom yes yes no 27 0 YFNERV+VnURWPalladioL Type 1 Custom yes yes no 32 0 GLCFZC+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 50 0 GOBEFB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 75 0 GOBEFC+PalatinoLinotype CID TrueType Identity-H yes yes yes 78 0 GOBEFD+PalatinoLinotype TrueType WinAnsi yes yes no 84 0 GOBEFF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 87 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 103 0 GOBEFB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 110 0 GOBEFD+PalatinoLinotype TrueType WinAnsi yes yes no 113 0 GOBEFF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 116 0 GOBEFB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 128 0 GOBEFD+PalatinoLinotype TrueType WinAnsi yes yes no 131 0 GOBEFF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 134 0 GOBEFB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 149 0 GOBEFD+PalatinoLinotype TrueType WinAnsi yes yes no 152 0 GOBEFF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 155 0 JHYTSG+CMR10 Type 1 Builtin yes yes no 164 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-09-14 02:29:37 CEST RepresentationInformation: pmc_10295744.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-09-13 02:17:58 CEST Size: 1605894 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 433 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Thymidine Analogues and Cell Toxicity Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Saturating S-Phase Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Estimating the CELL Cycle Time after a Single Administration of a Given Thymidine Analogue Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Inferring Duration and Phases of the Cell Cycle after Cumulative Labeling Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Estimating the Cell Cycle Time and Duration of the Synthetic Phase after Double Labeling Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Info: Title: Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues Author: Joaquín Martí-Clúa Subject: Tritiated thymidine autoradiography, 5-bromo-2'-deoxyuridine (BrdU) 5-chloro-2'-deoxyuridine (CldU), 5-iodo-2'-deoxyuridine (IdU), and 5-ethynyl-2'-deoxyiridine (EdU) labeling have been used for identifying the fraction of cells undergoing the S-phase of the cell cycle and to follow the fate of these cells during the embryonic, perinatal, and adult life in several species of vertebrate. In this current review, I will discuss the dosage and times of exposition to the aforementioned thymidine analogues to label most of the cells undergoing the S-phase of the cell cycle. I will also show how to infer, in an asynchronous cell population, the duration of the G1, S, and G2 phases, as well as the growth fraction and the span of the whole cell cycle on the base of some labeling schemes involving a single administration, continuous nucleotide analogue delivery, and double labeling with two thymidine analogues. In this context, the choice of the optimal dose of BrdU, CldU, IdU, and EdU to label S-phase cells is a pivotal aspect to produce neither cytotoxic effects nor alter cell cycle progression. I hope that the information presented in this review can be of use as a reference for researchers involved in the genesis of tissues and organs. 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