High throughput screening of small molecules for enhancing adipogenesis in vitro
Adipose tissues have been known to be the major site for energy storage and metabolic maintenance. Despite being researched for many decades, expanding and culturing primary adipocytes has remained difficult. To circumvent this problem, many labs in the field are using adipocytes differentiated from...
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| Other Authors: | |
| Format: | Final Year Project (FYP) |
| Language: | English |
| Published: |
2018
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| Online Access: | http://hdl.handle.net/10356/74179 |
| _version_ | 1826114237361029120 |
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| author | Goh, Jun Wei |
| author2 | Nuri Kim |
| author_facet | Nuri Kim Goh, Jun Wei |
| author_sort | Goh, Jun Wei |
| collection | NTU |
| description | Adipose tissues have been known to be the major site for energy storage and metabolic maintenance. Despite being researched for many decades, expanding and culturing primary adipocytes has remained difficult. To circumvent this problem, many labs in the field are using adipocytes differentiated from induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs), which can expand much more easily. However, these iPS-MSC-adipocytes are typically very immature, with only low levels of lipogenesis. In this study, we utilised small molecule high throughput screening (HTS) methods to promote the adipogenic differentiation and maturation of iPS-MSCs. We found that a small molecule combination 2+DO was able to significantly promote and enhance adipogenesis. We also optimized the iPS-MSC seeding density and glucose concentration to optimize subsequent adipogenesis. Finally, we tested our small molecule combination 2+DO on primary pre-adipocytes as well. RNA profiling by qRT- PCR showed that many adipogenic biomarkers such as LPL, SREBF1, FGF2, IRS2 and PPARG were significantly upregulated by 2+DO, as compared to normal adipogenic media. Taken together, we have demonstrated that HTS is a viable method for screening novel small molecules that enhance adipogenesis. This platform will provide a strong foundation to model fat-related metabolic diseases in vitro in future. |
| first_indexed | 2024-10-01T03:36:12Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/74179 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:36:12Z |
| publishDate | 2018 |
| record_format | dspace |
| spelling | ntu-10356/741792023-02-28T18:03:14Z High throughput screening of small molecules for enhancing adipogenesis in vitro Goh, Jun Wei Nuri Kim School of Biological Sciences A*STAR Genome Institute of Singapore Ng Shyh-Chang DRNTU::Science Adipose tissues have been known to be the major site for energy storage and metabolic maintenance. Despite being researched for many decades, expanding and culturing primary adipocytes has remained difficult. To circumvent this problem, many labs in the field are using adipocytes differentiated from induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs), which can expand much more easily. However, these iPS-MSC-adipocytes are typically very immature, with only low levels of lipogenesis. In this study, we utilised small molecule high throughput screening (HTS) methods to promote the adipogenic differentiation and maturation of iPS-MSCs. We found that a small molecule combination 2+DO was able to significantly promote and enhance adipogenesis. We also optimized the iPS-MSC seeding density and glucose concentration to optimize subsequent adipogenesis. Finally, we tested our small molecule combination 2+DO on primary pre-adipocytes as well. RNA profiling by qRT- PCR showed that many adipogenic biomarkers such as LPL, SREBF1, FGF2, IRS2 and PPARG were significantly upregulated by 2+DO, as compared to normal adipogenic media. Taken together, we have demonstrated that HTS is a viable method for screening novel small molecules that enhance adipogenesis. This platform will provide a strong foundation to model fat-related metabolic diseases in vitro in future. Bachelor of Science in Biological Sciences 2018-05-04T03:24:07Z 2018-05-04T03:24:07Z 2018 Final Year Project (FYP) http://hdl.handle.net/10356/74179 en Nanyang Technological University 42 p. application/pdf |
| spellingShingle | DRNTU::Science Goh, Jun Wei High throughput screening of small molecules for enhancing adipogenesis in vitro |
| title | High throughput screening of small molecules for enhancing adipogenesis in vitro |
| title_full | High throughput screening of small molecules for enhancing adipogenesis in vitro |
| title_fullStr | High throughput screening of small molecules for enhancing adipogenesis in vitro |
| title_full_unstemmed | High throughput screening of small molecules for enhancing adipogenesis in vitro |
| title_short | High throughput screening of small molecules for enhancing adipogenesis in vitro |
| title_sort | high throughput screening of small molecules for enhancing adipogenesis in vitro |
| topic | DRNTU::Science |
| url | http://hdl.handle.net/10356/74179 |
| work_keys_str_mv | AT gohjunwei highthroughputscreeningofsmallmoleculesforenhancingadipogenesisinvitro |