RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells
Abstract Background Human hair follicle mesenchymal stem cells (hHFMSCs) isolated from hair follicles possess multilineage differentiation potential. OCT4 is a gene critically associated with pluripotency properties. The cell morphology and adhesion of hHFMSCs significantly changed after transductio...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-10-01
|
| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-020-01976-1 |
| _version_ | 1818257155882483712 |
|---|---|
| author | Xiaozhen Yu Pengpeng Sun Xingang Huang Hua Chen Weiqing Huang Yingchun Ruan Weina Jiang Xiaohua Tan Zhijing Liu |
| author_facet | Xiaozhen Yu Pengpeng Sun Xingang Huang Hua Chen Weiqing Huang Yingchun Ruan Weina Jiang Xiaohua Tan Zhijing Liu |
| author_sort | Xiaozhen Yu |
| collection | DOAJ |
| description | Abstract Background Human hair follicle mesenchymal stem cells (hHFMSCs) isolated from hair follicles possess multilineage differentiation potential. OCT4 is a gene critically associated with pluripotency properties. The cell morphology and adhesion of hHFMSCs significantly changed after transduction of OCT4 and two subpopulations emerged, including adherent cells and floating cell. Floating cells cultured in hematopoietic induction medium and stimulated with erythropoetic growth factors could transdifferentiate into mature erythrocytes, whereas adherent cells formed negligible hematopoietic colonies. The aim of this study was to reveal the role of cell morphology and adhesion on erythropoiesis induced by OCT4 in hHFMSCs and to characterize the molecular mechanisms involved. Methods Floating cell was separated from adherent cell by centrifugation of the upper medium during cell culture. Cell size was observed through flow cytometry and cell adhesion was tested by disassociation and adhesion assays. RNA sequencing was performed to detect genome-wide transcriptomes and identify differentially expressed genes. GO enrichment analysis and KEGG pathway analysis were performed to analysis the functions and pathways enriched by differentially expressed genes. The expression of tight junction core members was verified by qPCR and Western blot. A regulatory network was constructed to figure out the relationship between cell adhesin, cytoskeleton, pluripotency, and hematopoiesis. Results The overexpression of OCT4 influenced the morphology and adhesion of hHFMSCs. Transcripts in floating cells and adherent cells are quite different. Data analysis showed that upregulated genes in floating cells were mainly related to pluripotency, germ layer development (including hematopoiesis lineage development), and downregulated genes were mainly related to cell adhesion, cell junctions, and the cytoskeleton. Most molecules of the tight junction (TJ) pathway were downregulated and molecular homeostasis of the TJ was disturbed, as CLDNs were disrupted, and JAMs and TJPs were upregulated. The dynamic expression of cell adhesion-related gene E-cadherin and cytoskeleton-related gene ACTN2 might cause different morphology and adhesion. Finally, a regulatory network centered to OCT4 was constructed, which elucidated that he TJ pathway critically bridges pluripotency and hematopoiesis in a TJP1-dependent way. Conclusions Regulations of cell morphology and adhesion via the TJ pathway conducted by OCT4 might modulate hematopoiesis in hHFMSCs, thus developing potential mechanism of erythropoiesis in vitro. |
| first_indexed | 2024-12-12T17:39:10Z |
| format | Article |
| id | doaj.art-ad4a946668574d7cbb464d7734c6541a |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-12T17:39:10Z |
| publishDate | 2020-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-ad4a946668574d7cbb464d7734c6541a2022-12-22T00:17:09ZengBMCStem Cell Research & Therapy1757-65122020-10-0111111610.1186/s13287-020-01976-1RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cellsXiaozhen Yu0Pengpeng Sun1Xingang Huang2Hua Chen3Weiqing Huang4Yingchun Ruan5Weina Jiang6Xiaohua Tan7Zhijing Liu8Department of Pathology, College of Basic Medical Sciences, Qingdao UniversityDepartment of Critical Care Medicine, Qingdao Center Hospital, affiliated with Qingdao UniversityDepartment of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao UniversityDepartment of Pathology, College of Basic Medical Sciences, Qingdao UniversityDepartment of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao UniversityDepartment of Pathology, College of Basic Medical Sciences, Qingdao UniversityDepartment of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao UniversityDepartment of Pathology, College of Basic Medical Sciences, Qingdao UniversityDepartment of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao UniversityAbstract Background Human hair follicle mesenchymal stem cells (hHFMSCs) isolated from hair follicles possess multilineage differentiation potential. OCT4 is a gene critically associated with pluripotency properties. The cell morphology and adhesion of hHFMSCs significantly changed after transduction of OCT4 and two subpopulations emerged, including adherent cells and floating cell. Floating cells cultured in hematopoietic induction medium and stimulated with erythropoetic growth factors could transdifferentiate into mature erythrocytes, whereas adherent cells formed negligible hematopoietic colonies. The aim of this study was to reveal the role of cell morphology and adhesion on erythropoiesis induced by OCT4 in hHFMSCs and to characterize the molecular mechanisms involved. Methods Floating cell was separated from adherent cell by centrifugation of the upper medium during cell culture. Cell size was observed through flow cytometry and cell adhesion was tested by disassociation and adhesion assays. RNA sequencing was performed to detect genome-wide transcriptomes and identify differentially expressed genes. GO enrichment analysis and KEGG pathway analysis were performed to analysis the functions and pathways enriched by differentially expressed genes. The expression of tight junction core members was verified by qPCR and Western blot. A regulatory network was constructed to figure out the relationship between cell adhesin, cytoskeleton, pluripotency, and hematopoiesis. Results The overexpression of OCT4 influenced the morphology and adhesion of hHFMSCs. Transcripts in floating cells and adherent cells are quite different. Data analysis showed that upregulated genes in floating cells were mainly related to pluripotency, germ layer development (including hematopoiesis lineage development), and downregulated genes were mainly related to cell adhesion, cell junctions, and the cytoskeleton. Most molecules of the tight junction (TJ) pathway were downregulated and molecular homeostasis of the TJ was disturbed, as CLDNs were disrupted, and JAMs and TJPs were upregulated. The dynamic expression of cell adhesion-related gene E-cadherin and cytoskeleton-related gene ACTN2 might cause different morphology and adhesion. Finally, a regulatory network centered to OCT4 was constructed, which elucidated that he TJ pathway critically bridges pluripotency and hematopoiesis in a TJP1-dependent way. Conclusions Regulations of cell morphology and adhesion via the TJ pathway conducted by OCT4 might modulate hematopoiesis in hHFMSCs, thus developing potential mechanism of erythropoiesis in vitro.http://link.springer.com/article/10.1186/s13287-020-01976-1Tight junction pathwayHuman hair follicle mesenchymal stem cellsOCT4Hematopoiesis |
| spellingShingle | Xiaozhen Yu Pengpeng Sun Xingang Huang Hua Chen Weiqing Huang Yingchun Ruan Weina Jiang Xiaohua Tan Zhijing Liu RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells Stem Cell Research & Therapy Tight junction pathway Human hair follicle mesenchymal stem cells OCT4 Hematopoiesis |
| title | RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells |
| title_full | RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells |
| title_fullStr | RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells |
| title_full_unstemmed | RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells |
| title_short | RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells |
| title_sort | rna seq reveals tight junction relevant erythropoietic fate induced by oct4 in human hair follicle mesenchymal stem cells |
| topic | Tight junction pathway Human hair follicle mesenchymal stem cells OCT4 Hematopoiesis |
| url | http://link.springer.com/article/10.1186/s13287-020-01976-1 |
| work_keys_str_mv | AT xiaozhenyu rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT pengpengsun rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT xinganghuang rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT huachen rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT weiqinghuang rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT yingchunruan rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT weinajiang rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT xiaohuatan rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells AT zhijingliu rnaseqrevealstightjunctionrelevanterythropoieticfateinducedbyoct4inhumanhairfolliclemesenchymalstemcells |