دورية أكاديمية
Cytokinesis-block micronucleus assay adapted for analyzing genomic instability of human mesenchymal stem cells.
| العنوان: | Cytokinesis-block micronucleus assay adapted for analyzing genomic instability of human mesenchymal stem cells. |
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| المؤلفون: | Cornélio DA; 1 Departamento de Biologia Celular e Genética, Centro de Biociências, Universidade Federal do Rio Grande do Norte , Natal, Brazil ., Tavares JC, Pimentel TV, Cavalcanti GB Jr, Batistuzzo de Medeiros SR |
| المصدر: | Stem cells and development [Stem Cells Dev] 2014 Apr 15; Vol. 23 (8), pp. 823-38. Date of Electronic Publication: 2014 Feb 04. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Mary Ann Liebert, Inc Country of Publication: United States NLM ID: 101197107 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1557-8534 (Electronic) Linking ISSN: 15473287 NLM ISO Abbreviation: Stem Cells Dev Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Larchmont, NY : Mary Ann Liebert, Inc., c2004- |
| مواضيع طبية MeSH: | Genomic Instability*, Cytokinesis/*drug effects , Mesenchymal Stem Cells/*physiology, Biomarkers/metabolism ; Cell Differentiation ; Cell Line, Tumor ; Cell Nucleus ; Cell Shape ; Cytochalasin B/pharmacology ; Female ; Humans ; Infant, Newborn ; Male ; Micronucleus Tests/methods ; Middle Aged ; Primary Cell Culture |
| مستخلص: | Human mesenchymal stem cells (hMSCs) are multipotent cells used in cell therapy research. One of the problems involving hMSCs is the possibility of genetic instability during in vitro expansion required to obtain a suitable number of cells for clinical applications. The cytokinesis-block micronucleus (CBMN) assay measures genetic instability by analyzing the presence of micronucleus (MN), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in binucleated cells. The present study describes modifications in the CBMN assay methodology to analyze genetic instability in hMSCs isolated from the umbilical vein and in vitro expanded. The best protocol to achieve binucleated hMSCs with preserved cytoplasm was as follows: cytochalasin B concentration (4.0 μg/mL), use of hypotonic treatment (3 min), and the fixative solution (9 methanol:1 acetic acid). These adaptations were reproduced in three hMSC primary cell cultures and also in XP4PA and A549 cell lines. The frequency of hMSCs treated with mitomycin-C presenting MN was lower than that with other nuclear alterations, indicating that the hMSCs contain mechanisms to avoid a high level of chromosomal breaks. However, a high frequency of cells with NPBs was detected and spontaneous anaphase bridges under normal hMSC in vitro culture were observed. Considering that anaphase bridges are characteristic alterations in tumor cells, the CBMN assay is indicated as an important tool associated with other genetic analyses in order to ensure the safe clinical use of hMSCs in cell therapy. |
| المشرفين على المادة: | 0 (Biomarkers) 3CHI920QS7 (Cytochalasin B) |
| تواريخ الأحداث: | Date Created: 20131217 Date Completed: 20141201 Latest Revision: 20220330 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1089/scd.2013.0383 |
| PMID: | 24328548 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1557-8534 |
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| DOI: | 10.1089/scd.2013.0383 |