Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells
The differentiation of human pluripotent stem cells (hPSCs) to neural stem cells (NSCs) is the key initial event in neurogenesis and is thought to be dependent on the family of Wnt growth factors, their receptors and signaling proteins. The delineation of the transcriptional pathways that mediate Wn...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7473 |
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| author | Bing-Chun Liu Fang-Yuan Liu Xin-Yue Gao Yang-Lin Chen Qiao-Qiao Meng Yong-Li Song Xi-He Li Si-Qin Bao |
| author_facet | Bing-Chun Liu Fang-Yuan Liu Xin-Yue Gao Yang-Lin Chen Qiao-Qiao Meng Yong-Li Song Xi-He Li Si-Qin Bao |
| author_sort | Bing-Chun Liu |
| collection | DOAJ |
| description | The differentiation of human pluripotent stem cells (hPSCs) to neural stem cells (NSCs) is the key initial event in neurogenesis and is thought to be dependent on the family of Wnt growth factors, their receptors and signaling proteins. The delineation of the transcriptional pathways that mediate Wnt-induced hPSCs to NSCs differentiation is vital for understanding the global genomic mechanisms of the development of NSCs and, potentially, the creation of new protocols in regenerative medicine. To understand the genomic mechanism of Wnt signaling during NSCs development, we treated hPSCs with Wnt activator (CHIR-99021) and leukemia inhibitory factor (LIF) in a chemically defined medium (N2B27) to induce NSCs, referred to as CLNSCs. The CLNSCs were subcultured for more than 40 passages in vitro; were positive for AP staining; expressed neural progenitor markers such as <i>NESTIN</i>, <i>PAX6</i>, <i>SOX2</i>, and <i>SOX1</i>; and were able to differentiate into three neural lineage cells: neurons, astrocytes, and oligodendrocytes in vitro. Our transcriptome analyses revealed that the Wnt and Hedgehog signaling pathways regulate hPSCs cell fate decisions for neural lineages and maintain the self-renewal of CLNSCs. One interesting network could be the deregulation of the Wnt/β-catenin signaling pathway in CLNSCs via the downregulation of c-MYC, which may promote exit from pluripotency and neural differentiation. The Wnt-induced spinal markers <i>HOXA1-4</i>, <i>HOXA7</i>, <i>HOXB1-4,</i> and <i>HOXC4</i> were increased, however, the brain markers <i>FOXG1</i> and <i>OTX2</i>, were absent in the CLNSCs, indicating that CLNSCs have partial spinal cord properties. Finally, a CLNSC simple culture condition, when applied to hPSCs, supports the generation of NSCs, and provides a new and efficient cell model with which to untangle the mechanisms during neurogenesis. |
| first_indexed | 2024-03-10T09:37:12Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:37:12Z |
| publishDate | 2021-07-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-50e29ed2ac394490a156eeceec6af5132023-11-22T03:59:07ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214747310.3390/ijms22147473Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem CellsBing-Chun Liu0Fang-Yuan Liu1Xin-Yue Gao2Yang-Lin Chen3Qiao-Qiao Meng4Yong-Li Song5Xi-He Li6Si-Qin Bao7The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot 010020, ChinaThe differentiation of human pluripotent stem cells (hPSCs) to neural stem cells (NSCs) is the key initial event in neurogenesis and is thought to be dependent on the family of Wnt growth factors, their receptors and signaling proteins. The delineation of the transcriptional pathways that mediate Wnt-induced hPSCs to NSCs differentiation is vital for understanding the global genomic mechanisms of the development of NSCs and, potentially, the creation of new protocols in regenerative medicine. To understand the genomic mechanism of Wnt signaling during NSCs development, we treated hPSCs with Wnt activator (CHIR-99021) and leukemia inhibitory factor (LIF) in a chemically defined medium (N2B27) to induce NSCs, referred to as CLNSCs. The CLNSCs were subcultured for more than 40 passages in vitro; were positive for AP staining; expressed neural progenitor markers such as <i>NESTIN</i>, <i>PAX6</i>, <i>SOX2</i>, and <i>SOX1</i>; and were able to differentiate into three neural lineage cells: neurons, astrocytes, and oligodendrocytes in vitro. Our transcriptome analyses revealed that the Wnt and Hedgehog signaling pathways regulate hPSCs cell fate decisions for neural lineages and maintain the self-renewal of CLNSCs. One interesting network could be the deregulation of the Wnt/β-catenin signaling pathway in CLNSCs via the downregulation of c-MYC, which may promote exit from pluripotency and neural differentiation. The Wnt-induced spinal markers <i>HOXA1-4</i>, <i>HOXA7</i>, <i>HOXB1-4,</i> and <i>HOXC4</i> were increased, however, the brain markers <i>FOXG1</i> and <i>OTX2</i>, were absent in the CLNSCs, indicating that CLNSCs have partial spinal cord properties. Finally, a CLNSC simple culture condition, when applied to hPSCs, supports the generation of NSCs, and provides a new and efficient cell model with which to untangle the mechanisms during neurogenesis.https://www.mdpi.com/1422-0067/22/14/7473human pluripotent stem cellsneural stem cellsWnt signalingspinal cord |
| spellingShingle | Bing-Chun Liu Fang-Yuan Liu Xin-Yue Gao Yang-Lin Chen Qiao-Qiao Meng Yong-Li Song Xi-He Li Si-Qin Bao Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells International Journal of Molecular Sciences human pluripotent stem cells neural stem cells Wnt signaling spinal cord |
| title | Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells |
| title_full | Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells |
| title_fullStr | Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells |
| title_full_unstemmed | Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells |
| title_short | Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells |
| title_sort | global transcriptional analyses of the wnt induced development of neural stem cells from human pluripotent stem cells |
| topic | human pluripotent stem cells neural stem cells Wnt signaling spinal cord |
| url | https://www.mdpi.com/1422-0067/22/14/7473 |
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