MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation
Background: Human umbilical cord blood (HUCB) is an acceptable and readily accessible source of stem cells. There is an ongoing interest in cord blood stem cell therapies; however, little is known about the possible unfavorable effects of laboratory modifications on the isolated HUCB cells. The invo...
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| Format: | Article |
| Language: | English |
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Shiraz University of Medical Sciences
2017-01-01
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| Series: | Iranian Journal of Medical Sciences |
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| Online Access: | http://ijms.sums.ac.ir/index.php/IJMS/article/view/2503 |
| _version_ | 1818955282205638656 |
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| author | Saeid Shahrabi Saeid Kaviani Masoud Soleimani Ali Akbar Pourfathollah Behnaz Bakhshandeh Saeideh Hajizamani Najmaldin Saki |
| author_facet | Saeid Shahrabi Saeid Kaviani Masoud Soleimani Ali Akbar Pourfathollah Behnaz Bakhshandeh Saeideh Hajizamani Najmaldin Saki |
| author_sort | Saeid Shahrabi |
| collection | DOAJ |
| description | Background: Human umbilical cord blood (HUCB) is an acceptable and readily accessible source of stem cells. There is an ongoing interest in cord blood stem cell therapies; however, little is known about the possible unfavorable effects of laboratory modifications on the isolated HUCB cells. The involvement of miRNAs in several biological processes has been shown. The aim of this study was to evaluate the possible changes in miRNA expression profiles in CD133+ hematopoietic cells after in vitro culture.
Materials and Methods: HUCBCD133+ hematopoietic stem cells were isolated by magnetic-activated cell sorting, and then the cells were counted using flow cytometry. The cells were divided into 2 groups. In the first group, RNA was extracted and the cells of the second group were cultured in vitro for 12 days and then these cells were used to assay miRNAs expression using real-time qPCR. Results: The results showed that the expression of 349 out of 1,151 screened miRNAs was upregulated following a 12-day in vitro culture of CD133+ cells, whereas the expression of 293 miRNAs was downregulated. In addition, the expression of 509 miRNAs was not significantly altered. Another in-silico analysis involving the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to the selected miRNAs was also conducted. Conclusion: Based on our results, the in vitro expansion of HUCB resulted in altered expression levels of miRNAs. This study provides information on the effects of 2-dimensional culture of hematopoietic cells prior to transplantation for more successful transplantation. |
| first_indexed | 2024-12-20T10:35:35Z |
| format | Article |
| id | doaj.art-92ed3d56f9d749f48d5a30aa1e976415 |
| institution | Directory Open Access Journal |
| issn | 0253-0716 1735-3688 |
| language | English |
| last_indexed | 2024-12-20T10:35:35Z |
| publishDate | 2017-01-01 |
| publisher | Shiraz University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Medical Sciences |
| spelling | doaj.art-92ed3d56f9d749f48d5a30aa1e9764152022-12-21T19:43:37ZengShiraz University of Medical SciencesIranian Journal of Medical Sciences0253-07161735-36882017-01-014214047MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to TransplantationSaeid Shahrabi0Saeid Kaviani1Masoud Soleimani2Ali Akbar Pourfathollah3Behnaz Bakhshandeh4Saeideh Hajizamani5Najmaldin Saki6Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran; and Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran; and Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Biotechnology, College of Science, University of Tehran, Tehran, IranDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, IranHealth Research Institute, Thalassemia and Hemoglobinopathy Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranBackground: Human umbilical cord blood (HUCB) is an acceptable and readily accessible source of stem cells. There is an ongoing interest in cord blood stem cell therapies; however, little is known about the possible unfavorable effects of laboratory modifications on the isolated HUCB cells. The involvement of miRNAs in several biological processes has been shown. The aim of this study was to evaluate the possible changes in miRNA expression profiles in CD133+ hematopoietic cells after in vitro culture. Materials and Methods: HUCBCD133+ hematopoietic stem cells were isolated by magnetic-activated cell sorting, and then the cells were counted using flow cytometry. The cells were divided into 2 groups. In the first group, RNA was extracted and the cells of the second group were cultured in vitro for 12 days and then these cells were used to assay miRNAs expression using real-time qPCR. Results: The results showed that the expression of 349 out of 1,151 screened miRNAs was upregulated following a 12-day in vitro culture of CD133+ cells, whereas the expression of 293 miRNAs was downregulated. In addition, the expression of 509 miRNAs was not significantly altered. Another in-silico analysis involving the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to the selected miRNAs was also conducted. Conclusion: Based on our results, the in vitro expansion of HUCB resulted in altered expression levels of miRNAs. This study provides information on the effects of 2-dimensional culture of hematopoietic cells prior to transplantation for more successful transplantation.http://ijms.sums.ac.ir/index.php/IJMS/article/view/2503Hematopoietic stem cellsUmbilical cord bloodMicroRNAsCD133Signal transduction |
| spellingShingle | Saeid Shahrabi Saeid Kaviani Masoud Soleimani Ali Akbar Pourfathollah Behnaz Bakhshandeh Saeideh Hajizamani Najmaldin Saki MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation Iranian Journal of Medical Sciences Hematopoietic stem cells Umbilical cord blood MicroRNAs CD133 Signal transduction |
| title | MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation |
| title_full | MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation |
| title_fullStr | MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation |
| title_full_unstemmed | MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation |
| title_short | MicroRNA Modulation during the In vitro Culture of Hematopoietic Stem Cells Prior to Transplantation |
| title_sort | microrna modulation during the in vitro culture of hematopoietic stem cells prior to transplantation |
| topic | Hematopoietic stem cells Umbilical cord blood MicroRNAs CD133 Signal transduction |
| url | http://ijms.sums.ac.ir/index.php/IJMS/article/view/2503 |
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