Whole transcriptome scanning and validation of negatively related genes in UC-MSCs
Background: Human umbilical cord mesenchymal stem cells (UC-MSCs) are one of the most extensively researched stem cell types due to their potential for multi-lineage differentiation, secretion of regenerative factors, modulations of immunological activities, and the release of regenerative substance...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024040271 |
| _version_ | 1797224120299028480 |
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| author | Linghan Tian Weibin Wang Xuzhen Li Yan Chen Qian Song Lu Yuan Tingting Hao Jiaming Gu Jian Dong |
| author_facet | Linghan Tian Weibin Wang Xuzhen Li Yan Chen Qian Song Lu Yuan Tingting Hao Jiaming Gu Jian Dong |
| author_sort | Linghan Tian |
| collection | DOAJ |
| description | Background: Human umbilical cord mesenchymal stem cells (UC-MSCs) are one of the most extensively researched stem cell types due to their potential for multi-lineage differentiation, secretion of regenerative factors, modulations of immunological activities, and the release of regenerative substances and influence immunological processes. Since UC-MSCs must be cultivated on a large scale for clinical use, selecting the appropriate storing passage, such as the usage-based passage of UC-MSCs, is critical for long-term autologous or allogeneic usage. Long-term cultivation of stem cells, on the other hand, causes them to lose their pluripotent differentiation capacity. As a result, distinguishing between high and low passages of UC-MSCs and identifying the particular variations associated with stem cells and their modes of action is essential for regenerative medicine. Therefore, we investigated the biological features and transcriptional changes of UC-MSCs over passages. Methods: UC-MSCs were isolated from the tissues of the human umbilical cord, and UC-MSCs from five passages (P1, P3, P5, P10 and P15) with three repetitions were compared and identified based on morphology, cell markers, differentiation capacity, and aging-related characteristics. It was previously assumed that the phenotype of cells before the P10 passage was stable, defined as early passage, and that culture could be continued until the 15th passage, defined as late passage. Next, the five passages of UC-MSCs were sequenced using high-throughput complete transcriptome sequencing. Fuzzy C-Means Clustering (FCM) and Weighted Gene Co-expression Network Analysis (WGCNA) were used to find hub genes, and gene silencing was performed to investigate the impact of missing genes on the stemness of UC-MSC cells. Results: UC-MSCs of different passages displayed similar surface markers, including CD73, CD105, CD90, CD34, CD45 and HLA-DR. However, the proliferation time of late-phase UC-MSCs was longer than that of early-phase UC-MSCs, and the expression of the senescence-associated (SA)-β-gal staining marker was higher. At the same time, pluripotency markers (NANOG, OCT4, SOX2 and KIF4A) were down-regulated, and the multi-differentiation potential was reduced. Meanwhile, KIFC1 and UBE2C were down-regulated in late-phase UC-MSCs, which were involved in the maintenance of stemness. Conclusions: KIFC1 and UBE2C were highly expressed in early-UC-MSCs and showed a downward gradient trend with cell expansion in vitro. They regulated UC-MSC proliferation, colony sphere formation, multiple differentiation, stemness maintenance, and other biological manifestations. Therefore, they are anticipated to be new biomarkers for UC-MSCs quality identification in regenerative medicine applications. |
| first_indexed | 2024-04-24T13:48:04Z |
| format | Article |
| id | doaj.art-404032b1a7f64bc0b99baceafece4baf |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-04-24T13:48:04Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-404032b1a7f64bc0b99baceafece4baf2024-04-04T05:06:35ZengElsevierHeliyon2405-84402024-03-01106e27996Whole transcriptome scanning and validation of negatively related genes in UC-MSCsLinghan Tian0Weibin Wang1Xuzhen Li2Yan Chen3Qian Song4Lu Yuan5Tingting Hao6Jiaming Gu7Jian Dong8Department of Yunnan Tumor Research Institute, Kunming, 650118, China; The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China; Yunnan Cancer Hospital, Kunming, 650118, ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, 650201, China; Yunnan Research Institute for Local Plateau Agriculture and Industry, Kunming, 650201, ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, 650201, China; Yunnan Research Institute for Local Plateau Agriculture and Industry, Kunming, 650201, ChinaDepartment of Yunnan Tumor Research Institute, Kunming, 650118, China; The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China; Yunnan Cancer Hospital, Kunming, 650118, ChinaDepartment of Yunnan Tumor Research Institute, Kunming, 650118, China; The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China; Yunnan Cancer Hospital, Kunming, 650118, ChinaDepartment of Yunnan Tumor Research Institute, Kunming, 650118, China; The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China; Yunnan Cancer Hospital, Kunming, 650118, ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, 650201, China; Yunnan Research Institute for Local Plateau Agriculture and Industry, Kunming, 650201, ChinaDepartment of Yunnan Tumor Research Institute, Kunming, 650118, China; The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China; Yunnan Cancer Hospital, Kunming, 650118, ChinaDepartment of Yunnan Tumor Research Institute, Kunming, 650118, China; The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China; Yunnan Cancer Hospital, Kunming, 650118, China; Corresponding author. The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118, China.Background: Human umbilical cord mesenchymal stem cells (UC-MSCs) are one of the most extensively researched stem cell types due to their potential for multi-lineage differentiation, secretion of regenerative factors, modulations of immunological activities, and the release of regenerative substances and influence immunological processes. Since UC-MSCs must be cultivated on a large scale for clinical use, selecting the appropriate storing passage, such as the usage-based passage of UC-MSCs, is critical for long-term autologous or allogeneic usage. Long-term cultivation of stem cells, on the other hand, causes them to lose their pluripotent differentiation capacity. As a result, distinguishing between high and low passages of UC-MSCs and identifying the particular variations associated with stem cells and their modes of action is essential for regenerative medicine. Therefore, we investigated the biological features and transcriptional changes of UC-MSCs over passages. Methods: UC-MSCs were isolated from the tissues of the human umbilical cord, and UC-MSCs from five passages (P1, P3, P5, P10 and P15) with three repetitions were compared and identified based on morphology, cell markers, differentiation capacity, and aging-related characteristics. It was previously assumed that the phenotype of cells before the P10 passage was stable, defined as early passage, and that culture could be continued until the 15th passage, defined as late passage. Next, the five passages of UC-MSCs were sequenced using high-throughput complete transcriptome sequencing. Fuzzy C-Means Clustering (FCM) and Weighted Gene Co-expression Network Analysis (WGCNA) were used to find hub genes, and gene silencing was performed to investigate the impact of missing genes on the stemness of UC-MSC cells. Results: UC-MSCs of different passages displayed similar surface markers, including CD73, CD105, CD90, CD34, CD45 and HLA-DR. However, the proliferation time of late-phase UC-MSCs was longer than that of early-phase UC-MSCs, and the expression of the senescence-associated (SA)-β-gal staining marker was higher. At the same time, pluripotency markers (NANOG, OCT4, SOX2 and KIF4A) were down-regulated, and the multi-differentiation potential was reduced. Meanwhile, KIFC1 and UBE2C were down-regulated in late-phase UC-MSCs, which were involved in the maintenance of stemness. Conclusions: KIFC1 and UBE2C were highly expressed in early-UC-MSCs and showed a downward gradient trend with cell expansion in vitro. They regulated UC-MSC proliferation, colony sphere formation, multiple differentiation, stemness maintenance, and other biological manifestations. Therefore, they are anticipated to be new biomarkers for UC-MSCs quality identification in regenerative medicine applications.http://www.sciencedirect.com/science/article/pii/S2405844024040271Stem cellsUC-MSCsDifferentiationStemnessUmbilical cord |
| spellingShingle | Linghan Tian Weibin Wang Xuzhen Li Yan Chen Qian Song Lu Yuan Tingting Hao Jiaming Gu Jian Dong Whole transcriptome scanning and validation of negatively related genes in UC-MSCs Heliyon Stem cells UC-MSCs Differentiation Stemness Umbilical cord |
| title | Whole transcriptome scanning and validation of negatively related genes in UC-MSCs |
| title_full | Whole transcriptome scanning and validation of negatively related genes in UC-MSCs |
| title_fullStr | Whole transcriptome scanning and validation of negatively related genes in UC-MSCs |
| title_full_unstemmed | Whole transcriptome scanning and validation of negatively related genes in UC-MSCs |
| title_short | Whole transcriptome scanning and validation of negatively related genes in UC-MSCs |
| title_sort | whole transcriptome scanning and validation of negatively related genes in uc mscs |
| topic | Stem cells UC-MSCs Differentiation Stemness Umbilical cord |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024040271 |
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