Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture
Human primary fibroblasts are a popular type of somatic cells for the production of induced pluripotent stem (iPS) cells. Here we characterized biological properties of primary fibroblasts in terms of cell-growth rate, cytogenetic stability, and the number of inactive X chromosomes during long-term...
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| Format: | Article |
| Language: | English |
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Wiley
2010-01-01
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| Series: | International Journal of Cell Biology |
| Online Access: | http://dx.doi.org/10.1155/2010/289653 |
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| author | Zhi-Gang Xue Zhan-Ping Shi Juan Dong Ting-Ting Liao Yan-Peng Wang Xue-Ping Sun Zheng-Jie Yan Xiao-Qiao Qian Yu-Gui Cui Juan Chen Jia-Yin Liu Guoping Fan |
| author_facet | Zhi-Gang Xue Zhan-Ping Shi Juan Dong Ting-Ting Liao Yan-Peng Wang Xue-Ping Sun Zheng-Jie Yan Xiao-Qiao Qian Yu-Gui Cui Juan Chen Jia-Yin Liu Guoping Fan |
| author_sort | Zhi-Gang Xue |
| collection | DOAJ |
| description | Human primary fibroblasts are a popular type of somatic cells for the production of induced pluripotent stem (iPS) cells. Here we characterized biological properties of primary fibroblasts in terms of cell-growth rate, cytogenetic stability, and the number of inactive X chromosomes during long-term passaging. We produced eight lines of female human dermal fibroblasts (HDFs) and found normal karyotype and expected pattern of X chromosome inactivation (XCI) at low passages (Passage P1-5). However, four out of the eight HDF lines at high passage numbers (≥P10) exhibited duplicated hallmarks of inactive X chromosome including two punctuate signals of histone H3 lysine 27 trimethylation (H3K27me3) and X inactive-specific transcript (XIST) RNA signals in approximately 8.5–18.5% of the cells. Our data suggest that the copy number of inactive X chromosomes in a subset of female HDF is increased by a two-fold. Consistently, DNA fluorescent in situ hybridization (FISH) identified 3-4 copies of X chromosomes in one nucleus in this subset of cells with two inactive Xs. We conclude that female HDF cultures exhibit a higher risk of genetic anomalies such as carrying an increased number of X chromosomes including both active and inactive X chromosomes at a high passage (≥P10). |
| format | Article |
| id | doaj-art-a9e82d463d3041a9a9675ca1a2fc67a0 |
| institution | Kabale University |
| issn | 1687-8876 1687-8884 |
| language | English |
| publishDate | 2010-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Cell Biology |
| spelling | doaj-art-a9e82d463d3041a9a9675ca1a2fc67a02025-02-03T05:44:04ZengWileyInternational Journal of Cell Biology1687-88761687-88842010-01-01201010.1155/2010/289653289653Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term CultureZhi-Gang Xue0Zhan-Ping Shi1Juan Dong2Ting-Ting Liao3Yan-Peng Wang4Xue-Ping Sun5Zheng-Jie Yan6Xiao-Qiao Qian7Yu-Gui Cui8Juan Chen9Jia-Yin Liu10Guoping Fan11Jiangsu Province Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, ChinaJiangsu Province Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCenter of Clinical Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaJiangsu Province Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, ChinaJiangsu Province Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, ChinaDepartment of Human Genetics, University of California Los Angeles, Los Angeles, CA 90095, USAHuman primary fibroblasts are a popular type of somatic cells for the production of induced pluripotent stem (iPS) cells. Here we characterized biological properties of primary fibroblasts in terms of cell-growth rate, cytogenetic stability, and the number of inactive X chromosomes during long-term passaging. We produced eight lines of female human dermal fibroblasts (HDFs) and found normal karyotype and expected pattern of X chromosome inactivation (XCI) at low passages (Passage P1-5). However, four out of the eight HDF lines at high passage numbers (≥P10) exhibited duplicated hallmarks of inactive X chromosome including two punctuate signals of histone H3 lysine 27 trimethylation (H3K27me3) and X inactive-specific transcript (XIST) RNA signals in approximately 8.5–18.5% of the cells. Our data suggest that the copy number of inactive X chromosomes in a subset of female HDF is increased by a two-fold. Consistently, DNA fluorescent in situ hybridization (FISH) identified 3-4 copies of X chromosomes in one nucleus in this subset of cells with two inactive Xs. We conclude that female HDF cultures exhibit a higher risk of genetic anomalies such as carrying an increased number of X chromosomes including both active and inactive X chromosomes at a high passage (≥P10).http://dx.doi.org/10.1155/2010/289653 |
| spellingShingle | Zhi-Gang Xue Zhan-Ping Shi Juan Dong Ting-Ting Liao Yan-Peng Wang Xue-Ping Sun Zheng-Jie Yan Xiao-Qiao Qian Yu-Gui Cui Juan Chen Jia-Yin Liu Guoping Fan Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture International Journal of Cell Biology |
| title | Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture |
| title_full | Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture |
| title_fullStr | Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture |
| title_full_unstemmed | Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture |
| title_short | Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture |
| title_sort | evaluation of x inactivation status and cytogenetic stability of human dermal fibroblasts after long term culture |
| url | http://dx.doi.org/10.1155/2010/289653 |
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