Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study
Abstract Background Hepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) hold great promise in toxicological applications as well as in regenerative medicine. Previous efforts on hepatocyte differentiation have mostly relied on the use of growth factors (GFs) to rec...
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| Format: | Article |
| Language: | English |
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BMC
2020-09-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | http://link.springer.com/article/10.1186/s13287-020-01914-1 |
| _version_ | 1818359253905178624 |
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| author | Xiugong Gao Rong Li Patrick Cahan Yang Zhao Jeffrey J. Yourick Robert L. Sprando |
| author_facet | Xiugong Gao Rong Li Patrick Cahan Yang Zhao Jeffrey J. Yourick Robert L. Sprando |
| author_sort | Xiugong Gao |
| collection | DOAJ |
| description | Abstract Background Hepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) hold great promise in toxicological applications as well as in regenerative medicine. Previous efforts on hepatocyte differentiation have mostly relied on the use of growth factors (GFs) to recapitulate developmental signals under in vitro conditions. Recently, the use of small molecules (SMs) has emerged as an attractive tool to induce cell fate transition due to its superiority in terms of both quality and cost. However, HLCs derived using SMs have not been well characterized, especially on the transcriptome level. Methods HLCs were differentiated from human iPSCs using a protocol that only involves SMs and characterized by transcriptomic analysis using whole genome microarrays. Results HLCs derived using the SM protocol (HLC_SM) displayed specific hepatic marker expression and demonstrated key hepatic functions. Transcriptomic analysis of the SM-driven differentiation defined a hepatocyte differentiation track and characterized the expression of some key marker genes in major stages of hepatocyte differentiation. In addition, HLC_SM were scored with CellNet, a bioinformatics tool quantifying how closely engineered cell populations resemble their target cell type, and compared to primary human hepatocytes (PHHs), adult liver tissue, fetal liver tissue, HLCs differentiated using GFs (HLC_GF), and commercially available HLCs. Similar to HLC_GF, HLC_SM displayed a mixed phenotype of fetal and adult hepatocytes and had relatively low expression of metabolic enzymes, transporters, and nuclear receptors compared to PHHs. Finally, the differentially expressed genes in HLC_SM compared to HLC_GF and to PHHs were analyzed to identify pathways and upstream transcription regulators which could potentially be manipulated to improve the differentiation of HLCs. Conclusions Overall, the present study demonstrated the usefulness of the SM-based hepatocyte differentiation method, offered new insights into the molecular basis of hepatogenesis and associated gene regulation, and suggested ways for further improvements in hepatocyte differentiation in order to obtain more mature HLCs that could be used in toxicological studies. |
| first_indexed | 2024-12-13T20:41:58Z |
| format | Article |
| id | doaj.art-050a29d497e046bcbda45ab772adde80 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-13T20:41:58Z |
| publishDate | 2020-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-050a29d497e046bcbda45ab772adde802022-12-21T23:32:07ZengBMCStem Cell Research & Therapy1757-65122020-09-0111112110.1186/s13287-020-01914-1Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic studyXiugong Gao0Rong Li1Patrick Cahan2Yang Zhao3Jeffrey J. Yourick4Robert L. Sprando5Division of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug AdministrationDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug AdministrationDepartment of Biomedical Engineering, Institute for Cell Engineering, Johns Hopkins University School of MedicineDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug AdministrationDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug AdministrationDivision of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug AdministrationAbstract Background Hepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) hold great promise in toxicological applications as well as in regenerative medicine. Previous efforts on hepatocyte differentiation have mostly relied on the use of growth factors (GFs) to recapitulate developmental signals under in vitro conditions. Recently, the use of small molecules (SMs) has emerged as an attractive tool to induce cell fate transition due to its superiority in terms of both quality and cost. However, HLCs derived using SMs have not been well characterized, especially on the transcriptome level. Methods HLCs were differentiated from human iPSCs using a protocol that only involves SMs and characterized by transcriptomic analysis using whole genome microarrays. Results HLCs derived using the SM protocol (HLC_SM) displayed specific hepatic marker expression and demonstrated key hepatic functions. Transcriptomic analysis of the SM-driven differentiation defined a hepatocyte differentiation track and characterized the expression of some key marker genes in major stages of hepatocyte differentiation. In addition, HLC_SM were scored with CellNet, a bioinformatics tool quantifying how closely engineered cell populations resemble their target cell type, and compared to primary human hepatocytes (PHHs), adult liver tissue, fetal liver tissue, HLCs differentiated using GFs (HLC_GF), and commercially available HLCs. Similar to HLC_GF, HLC_SM displayed a mixed phenotype of fetal and adult hepatocytes and had relatively low expression of metabolic enzymes, transporters, and nuclear receptors compared to PHHs. Finally, the differentially expressed genes in HLC_SM compared to HLC_GF and to PHHs were analyzed to identify pathways and upstream transcription regulators which could potentially be manipulated to improve the differentiation of HLCs. Conclusions Overall, the present study demonstrated the usefulness of the SM-based hepatocyte differentiation method, offered new insights into the molecular basis of hepatogenesis and associated gene regulation, and suggested ways for further improvements in hepatocyte differentiation in order to obtain more mature HLCs that could be used in toxicological studies.http://link.springer.com/article/10.1186/s13287-020-01914-1Hepatocyte differentiationHepatocyte-like cellsInduced pluripotent stem cellsTranscriptomicsSmall moleculesMicroarray |
| spellingShingle | Xiugong Gao Rong Li Patrick Cahan Yang Zhao Jeffrey J. Yourick Robert L. Sprando Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study Stem Cell Research & Therapy Hepatocyte differentiation Hepatocyte-like cells Induced pluripotent stem cells Transcriptomics Small molecules Microarray |
| title | Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study |
| title_full | Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study |
| title_fullStr | Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study |
| title_full_unstemmed | Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study |
| title_short | Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study |
| title_sort | hepatocyte like cells derived from human induced pluripotent stem cells using small molecules implications of a transcriptomic study |
| topic | Hepatocyte differentiation Hepatocyte-like cells Induced pluripotent stem cells Transcriptomics Small molecules Microarray |
| url | http://link.springer.com/article/10.1186/s13287-020-01914-1 |
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