Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells
Background: Telomerase as an enzyme with reverse transcriptase activity has an essential role in telomere maintenance by adding a telomere repeat sequence to the 3' end of chromosome and is important for regulating of many processes in embryonic development including cell proliferation and diff...
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| Format: | Article |
| Language: | fas |
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Tehran University of Medical Sciences
2016-08-01
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| Series: | Tehran University Medical Journal |
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| Online Access: | http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5472&slc_lang=en&sid=1 |
| _version_ | 1818652710226886656 |
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| author | Leila Hosseinzadeh Anvar Saeid Hosseini-Asl Mohsen Sagha |
| author_facet | Leila Hosseinzadeh Anvar Saeid Hosseini-Asl Mohsen Sagha |
| author_sort | Leila Hosseinzadeh Anvar |
| collection | DOAJ |
| description | Background: Telomerase as an enzyme with reverse transcriptase activity has an essential role in telomere maintenance by adding a telomere repeat sequence to the 3' end of chromosome and is important for regulating of many processes in embryonic development including cell proliferation and differentiation. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) with a self-renewal capacity are cells that can differentiate into various germ layer derivatives including neural cells and cardiomyocytes, and undergo biological changes during long-term cultivation. Hence, the passage number in which the cells expanded seems to be very important for proliferating and differentiating. This study was aimed at investigating the relationship between the telomerase activity and the growth rate of (hUC-MSCs) at different passages.
Methods: This experimental study was performed in Ardabil University of Medical Sciences, Iran, from March 2014 to December 2014. The umbilical cord samples were obtained from full-term neonate hospitalized in Alavi’s Hospital in Ardabil under sterile conditions. The umbilical vessels were clear off and the small pieces of the umbilical cord were cultured in Dulbecco's modified eagle's medium (DMEM) supplemented with 20% fetal bovine serum (FBS). Then, the hUC-MSCs were harvested from passage one to three to calculate the population doubling time (PDT) and extract proteins by using CHAPS lysis buffer. Finally, the telomerase activity of the cells at different passages was measured by telomeric repeat amplification protocol (TRAP) and qRT-TRAP assays.
Results: The hUC-MSCs population doubling time at passage from 1 to 3 were calculated as the average of 54.68±1.92, 55.03±1.71 and 69.41±2.54 hours, respectively, suggesting the higher cell passage number, the more extended PDT. The threshold cycles (CTs) for the telomerase activity also showed 30.58±0.51, 27.24±0.74 and 32.13±0.75 for the cell passage from one to three, respectively, representing the significant increasing in telomerase activity at passage two compared with the other passages (P= 0.021).
Conclusion: Analysis of the growth curve, PDT determination and measurement of telomerase activity of the human umbilical cord-derived mesenchymal stem cells showed that the long-term cell culture can affect on the cell proliferation and the telomerase activity. |
| first_indexed | 2024-12-17T02:26:20Z |
| format | Article |
| id | doaj.art-427ebaceb731468eaa5a120294364328 |
| institution | Directory Open Access Journal |
| issn | 1683-1764 1735-7322 |
| language | fas |
| last_indexed | 2024-12-17T02:26:20Z |
| publishDate | 2016-08-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Tehran University Medical Journal |
| spelling | doaj.art-427ebaceb731468eaa5a1202943643282022-12-21T22:07:06ZfasTehran University of Medical SciencesTehran University Medical Journal1683-17641735-73222016-08-01745321329Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cellsLeila Hosseinzadeh Anvar0Saeid Hosseini-Asl1Mohsen Sagha2 Department of Biology, Islamic Azad University, Ahar Branch, Ahar, Iran. Department of Genetics, Laboratory of Medical Genetics, Imam Khomeini's Hospital, Ardabil University of Medical Sciences, Ardabil, Iran. Department of Genetics, Laboratory of Medical Genetics, Imam Khomeini's Hospital, Ardabil University of Medical Sciences, Ardabil, Iran. Department of Anatomical Sciences and Pathology, Research Laboratory for Embryology and Stem Cells, Faculty of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran. Background: Telomerase as an enzyme with reverse transcriptase activity has an essential role in telomere maintenance by adding a telomere repeat sequence to the 3' end of chromosome and is important for regulating of many processes in embryonic development including cell proliferation and differentiation. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) with a self-renewal capacity are cells that can differentiate into various germ layer derivatives including neural cells and cardiomyocytes, and undergo biological changes during long-term cultivation. Hence, the passage number in which the cells expanded seems to be very important for proliferating and differentiating. This study was aimed at investigating the relationship between the telomerase activity and the growth rate of (hUC-MSCs) at different passages. Methods: This experimental study was performed in Ardabil University of Medical Sciences, Iran, from March 2014 to December 2014. The umbilical cord samples were obtained from full-term neonate hospitalized in Alavi’s Hospital in Ardabil under sterile conditions. The umbilical vessels were clear off and the small pieces of the umbilical cord were cultured in Dulbecco's modified eagle's medium (DMEM) supplemented with 20% fetal bovine serum (FBS). Then, the hUC-MSCs were harvested from passage one to three to calculate the population doubling time (PDT) and extract proteins by using CHAPS lysis buffer. Finally, the telomerase activity of the cells at different passages was measured by telomeric repeat amplification protocol (TRAP) and qRT-TRAP assays. Results: The hUC-MSCs population doubling time at passage from 1 to 3 were calculated as the average of 54.68±1.92, 55.03±1.71 and 69.41±2.54 hours, respectively, suggesting the higher cell passage number, the more extended PDT. The threshold cycles (CTs) for the telomerase activity also showed 30.58±0.51, 27.24±0.74 and 32.13±0.75 for the cell passage from one to three, respectively, representing the significant increasing in telomerase activity at passage two compared with the other passages (P= 0.021). Conclusion: Analysis of the growth curve, PDT determination and measurement of telomerase activity of the human umbilical cord-derived mesenchymal stem cells showed that the long-term cell culture can affect on the cell proliferation and the telomerase activity.http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5472&slc_lang=en&sid=1cell growth curve human umbilical cord-derivedmesenchymal stem cells telomerase activity |
| spellingShingle | Leila Hosseinzadeh Anvar Saeid Hosseini-Asl Mohsen Sagha Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells Tehran University Medical Journal cell growth curve human umbilical cord-derived mesenchymal stem cells telomerase activity |
| title | Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells |
| title_full | Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells |
| title_fullStr | Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells |
| title_full_unstemmed | Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells |
| title_short | Relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells |
| title_sort | relationship between the telomerase activity and the growth kinetics of the human umbilical cord derived mesenchymal stem cells |
| topic | cell growth curve human umbilical cord-derived mesenchymal stem cells telomerase activity |
| url | http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5472&slc_lang=en&sid=1 |
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