5b412f1a5d75a87115cf71c4980d9402 308592.pdf f854f2b2230b95956604c73343f2ea19f5d7061a 308592.pdf 118684a1cd30abf68cb3f627ee09039c8e90b885b206c4f8cfd7cb50d5e50f1b 308592.pdf Title: Impact of Overnight Storage of Human Atrial Myocytes on Intracellular Calcium Homeostasis and Electrophysiological Utility Subject: Human atrial myocytes afford an attractive experimental model to investigate mechanisms underlying electrophysiological alterations in cardiovascular disease. However, this model presents limitations, such as the availability of human atrial tissue and a variable yield of myocytes isolation. Therefore, we aimed to determine whether overnight storage can increase the time window where the electrophysiological properties of human atrial myocytes can be determined. To address this issue, human atrial myocytes isolated from patients undergoing cardiac surgery were used for patch-clamp experiments on the day of cell isolation (Day 1) and the following day (Day 2). The shape of the current–voltage (I–V) relationship for the calcium current (ICa) depended on the access resistance and the cell capacitance, with large cells (>75 pF) requiring a lower access resistance (<15 M) than small cells (<40 pF) to avoid distortion of the I–V curve. Importantly, overnight storage did not significantly affect (1) the ICa amplitude or properties, (2) sarcoplasmic reticulum calcium homeostasis or (3) the frequency-dependency of the beat-to-beat response. In conclusion, overnight storage of isolated human atrial myocytes at 4 C does not affect essential features of intracellular calcium homeostasis and, therefore, affords a simple protocol to extend the experimental lifetime of human atrial myocytes. Keywords: human atrial myocyte; patch-clamp technique; calcium homeostasis; sarcoplasmic reticulum; calcium currents; beat-to-beat response Author: Cristina Aceituno, David Revuelta, Verónica Jiménez-Sábado, Antonino Ginel, Cristina E. Molina and Leif Hove-Madsen Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Thu Nov 7 07:12:10 2024 CET ModDate: Thu Nov 7 07:16:08 2024 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2768502 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZZTQUO+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 LUJAXH+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 UADAXL+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 QLTUPL+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 SPXLNH+PazoMath Type 1 Builtin yes yes yes 32 0 WTAUCZ+CMSY10 Type 1 Builtin yes yes yes 37 0 OPZDHV+VnURWPalladioL Type 1 Custom yes yes yes 42 0 OVNEXW+EURM10 Type 1 Builtin yes yes yes 72 0 IPPNBH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 90 0 IPPNCI+PalatinoLinotype CID TrueType Identity-H yes yes yes 93 0 IPPNCJ+PalatinoLinotype TrueType WinAnsi yes yes no 99 0 IPPNCL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 102 0 IPPNGK+PalatinoLinotype TrueType MacRoman yes yes no 105 0 IPPNBH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 120 0 IPPNCI+PalatinoLinotype CID TrueType Identity-H yes yes yes 123 0 IPPNCJ+PalatinoLinotype TrueType WinAnsi yes yes no 129 0 IPPNCL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 132 0 IPPNGK+PalatinoLinotype TrueType MacRoman yes yes no 135 0 IPPNBH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 149 0 IPPNCI+PalatinoLinotype CID TrueType Identity-H yes yes yes 152 0 IPPNCJ+PalatinoLinotype TrueType WinAnsi yes yes no 158 0 IPPNCL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 161 0 IPPNGK+PalatinoLinotype TrueType MacRoman yes yes no 164 0 IPPNBH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 179 0 IPPNCI+PalatinoLinotype CID TrueType Identity-H yes yes yes 182 0 IPPNCJ+PalatinoLinotype TrueType WinAnsi yes yes no 188 0 IPPNCL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 191 0 IPPNGK+PalatinoLinotype TrueType MacRoman yes yes no 194 0 IPPNBH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 206 0 IPPNCI+PalatinoLinotype CID TrueType Identity-H yes yes yes 209 0 IPPNCJ+PalatinoLinotype TrueType WinAnsi yes yes no 215 0 IPPNCL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 218 0 IPPNGK+PalatinoLinotype TrueType MacRoman yes yes no 221 0 IPPNBH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 236 0 IPPNCI+PalatinoLinotype CID TrueType Identity-H yes yes yes 239 0 IPPNCJ+PalatinoLinotype TrueType WinAnsi yes yes no 245 0 IPPNCL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 248 0 IPPNGK+PalatinoLinotype TrueType MacRoman yes yes no 251 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-03-01 02:49:34 CET RepresentationInformation: 308592.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-02-28 12:53:03 CET Size: 2768502 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 401 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Item: Title: Results Destination: section.3 Children: Item: Title: Impact of Overnight Storage on L-Type Calcium Current Properties Destination: subsection.3.1 Item: Title: Impact on Sarcoplasmic Reticulum Calcium Homeostasis Destination: subsection.3.2 Item: Title: Impact on the Frequency-Dependent Beat-to-Beat Response Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Children: Item: Title: Main Findings Destination: subsection.4.1 Item: Title: Perforated Patch Recordings Destination: subsection.4.2 Item: Title: Overnight Storage of Human Atrial Myocytes Destination: subsection.4.3 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Info: Title: Impact of Overnight Storage of Human Atrial Myocytes on Intracellular Calcium Homeostasis and Electrophysiological Utility Author: Cristina Aceituno, David Revuelta, Verónica Jiménez-Sábado, Antonino Ginel, Cristina E. Molina and Leif Hove-Madsen Subject: Human atrial myocytes afford an attractive experimental model to investigate mechanisms underlying electrophysiological alterations in cardiovascular disease. However, this model presents limitations, such as the availability of human atrial tissue and a variable yield of myocytes isolation. Therefore, we aimed to determine whether overnight storage can increase the time window where the electrophysiological properties of human atrial myocytes can be determined. To address this issue, human atrial myocytes isolated from patients undergoing cardiac surgery were used for patch-clamp experiments on the day of cell isolation (Day 1) and the following day (Day 2). The shape of the current voltage (I V) relationship for the calcium current (ICa) depended on the access resistance and the cell capacitance, with large cells (>75 pF) requiring a lower access resistance (<15 M) than small cells (<40 pF) to avoid distortion of the I V curve. Importantly, overnight storage did not significantly affect (1) the ICa amplitude or properties, (2) sarcoplasmic reticulum calcium homeostasis or (3) the frequency-dependency of the beat-to-beat response. In conclusion, overnight storage of isolated human atrial myocytes at 4 C does not affect essential features of intracellular calcium homeostasis and, therefore, affords a simple protocol to extend the experimental lifetime of human atrial myocytes. 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