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Hepatocyte differentiation from human ES cells using the simple embryoid body formation method and the staged-additional cocktail
To induce hepatocytes from human embryonic stem (hES) cells easily and effectively, a simple suspension culture method that separates ES colonies with a scraper and transfers them into newly developed, nonadherent MPC (2-methacryloyloxyethyl phosphorylcholine) plates, and the staged-additional cockt...
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| Published in: | TheScientificWorld 2009-01, Vol.9, p.884-890 |
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| creator | Sasaki, Katsunori Ichikawa, Hinako Takei, Shunsuke No, Hee Sung Tomotsune, Daihachiro Kano, Yoshiya Yokoyama, Tadayuki Sirasawa, Sakiko Mogi, Akimi Yoshie, Susumu Sasaki, Shujiro Yamada, Satoshi Matsumoto, Ken Mizuguchi, Masahiro Yue, Fengming Tanaka, Yoshiki |
| description | To induce hepatocytes from human embryonic stem (hES) cells easily and effectively, a simple suspension culture method that separates ES colonies with a scraper and transfers them into newly developed, nonadherent MPC (2-methacryloyloxyethyl phosphorylcholine) plates, and the staged-additional cocktail method, including growth factors, cytokines, and Lanford serum-free medium, were developed and evaluated mainly by morphological analysis. The formed embryoid bodies (EBs) showed compact cellular agglomeration until day 4 and later formed coeloms in their interior. RT-PCR (reverse transcriptase-polymerase chain reaction) analysis showed that they are gene markers of the three germ layers. Mesenchymal cells with rough endoplasmic reticulum (rER) and extracellular matrix (ECM), and without junctions, were recognized in the interior of the EBs by transmission electron microscopy (TEM) in addition to epithelial cells. When they were stimulated by the staged-additional cocktail, they expressed albumin-positive immunoreactivity, indocyanine green (ICG) uptake, and typical ultrastructures of the hepatocytes, including bile canaliculi. These results indicate that these combined methods promote EB formation and hepatocyte differentiation from hES cells. |
| doi_str_mv | 10.1100/tsw.2009.97 |
| format | article |
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The formed embryoid bodies (EBs) showed compact cellular agglomeration until day 4 and later formed coeloms in their interior. RT-PCR (reverse transcriptase-polymerase chain reaction) analysis showed that they are gene markers of the three germ layers. Mesenchymal cells with rough endoplasmic reticulum (rER) and extracellular matrix (ECM), and without junctions, were recognized in the interior of the EBs by transmission electron microscopy (TEM) in addition to epithelial cells. When they were stimulated by the staged-additional cocktail, they expressed albumin-positive immunoreactivity, indocyanine green (ICG) uptake, and typical ultrastructures of the hepatocytes, including bile canaliculi. These results indicate that these combined methods promote EB formation and hepatocyte differentiation from hES cells.</description><identifier>ISSN: 1537-744X</identifier><identifier>ISSN: 2356-6140</identifier><identifier>EISSN: 1537-744X</identifier><identifier>DOI: 10.1100/tsw.2009.97</identifier><identifier>PMID: 19734962</identifier><language>eng</language><publisher>United States: John Wiley & Sons, Inc</publisher><subject>Base Sequence ; Cell differentiation ; Cell Line ; DNA Primers ; Embryonic development ; Embryonic stem cells ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - ultrastructure ; Health aspects ; Hepatocytes - cytology ; Humans ; Liver cells ; Microscopy, Electron, Transmission ; Physiological aspects ; Reverse Transcriptase Polymerase Chain Reaction ; Short Communication</subject><ispartof>TheScientificWorld, 2009-01, Vol.9, p.884-890</ispartof><rights>COPYRIGHT 2009 John Wiley & Sons, Inc.</rights><rights>Copyright © 2009 Katsunori Sasaki et al. 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c622t-5b72b342d6ad3430586c0625a20d95f94ad96f9620431fb30fc4ae0fb9e1f8f33</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5823185/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5823185/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19734962$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sasaki, Katsunori</creatorcontrib><creatorcontrib>Ichikawa, Hinako</creatorcontrib><creatorcontrib>Takei, Shunsuke</creatorcontrib><creatorcontrib>No, Hee Sung</creatorcontrib><creatorcontrib>Tomotsune, Daihachiro</creatorcontrib><creatorcontrib>Kano, Yoshiya</creatorcontrib><creatorcontrib>Yokoyama, Tadayuki</creatorcontrib><creatorcontrib>Sirasawa, Sakiko</creatorcontrib><creatorcontrib>Mogi, Akimi</creatorcontrib><creatorcontrib>Yoshie, Susumu</creatorcontrib><creatorcontrib>Sasaki, Shujiro</creatorcontrib><creatorcontrib>Yamada, Satoshi</creatorcontrib><creatorcontrib>Matsumoto, Ken</creatorcontrib><creatorcontrib>Mizuguchi, Masahiro</creatorcontrib><creatorcontrib>Yue, Fengming</creatorcontrib><creatorcontrib>Tanaka, Yoshiki</creatorcontrib><title>Hepatocyte differentiation from human ES cells using the simple embryoid body formation method and the staged-additional cocktail</title><title>TheScientificWorld</title><addtitle>ScientificWorldJournal</addtitle><description>To induce hepatocytes from human embryonic stem (hES) cells easily and effectively, a simple suspension culture method that separates ES colonies with a scraper and transfers them into newly developed, nonadherent MPC (2-methacryloyloxyethyl phosphorylcholine) plates, and the staged-additional cocktail method, including growth factors, cytokines, and Lanford serum-free medium, were developed and evaluated mainly by morphological analysis. The formed embryoid bodies (EBs) showed compact cellular agglomeration until day 4 and later formed coeloms in their interior. RT-PCR (reverse transcriptase-polymerase chain reaction) analysis showed that they are gene markers of the three germ layers. Mesenchymal cells with rough endoplasmic reticulum (rER) and extracellular matrix (ECM), and without junctions, were recognized in the interior of the EBs by transmission electron microscopy (TEM) in addition to epithelial cells. When they were stimulated by the staged-additional cocktail, they expressed albumin-positive immunoreactivity, indocyanine green (ICG) uptake, and typical ultrastructures of the hepatocytes, including bile canaliculi. These results indicate that these combined methods promote EB formation and hepatocyte differentiation from hES cells.</description><subject>Base Sequence</subject><subject>Cell differentiation</subject><subject>Cell Line</subject><subject>DNA Primers</subject><subject>Embryonic development</subject><subject>Embryonic stem cells</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - ultrastructure</subject><subject>Health aspects</subject><subject>Hepatocytes - cytology</subject><subject>Humans</subject><subject>Liver cells</subject><subject>Microscopy, Electron, Transmission</subject><subject>Physiological aspects</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Short Communication</subject><issn>1537-744X</issn><issn>2356-6140</issn><issn>1537-744X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNptksuLFDEQxhtR3IeevEtA8LL0mFenJxdhWVZ3YcGDCt5CdR49Wbs7Y5JZmaP_uWl70B2QHBKqfvWlqviq6hXBK0IwfpfTzxXFWK5k-6Q6JQ1r65bzb08fvU-qs5TuMWbrljTPqxMiW8aloKfVrxu7hRz0PltkvHM22il7yD5MyMUwos1uhAldf0baDkNCu-SnHuWNRcmP28EiO3ZxH7xBXTB75EIcl-LR5k0wCCaz0Bl6a2owxs9pGJAO-nsGP7yonjkYkn15uM-rrx-uv1zd1HefPt5eXd7VWlCa66Zracc4NQIM4ww3a6GxoA1QbGTjJAcjhSszYc6I6xh2moPFrpOWuLVj7Ly6XXRNgHu1jX6EuFcBvPoTCLFXELPXg1VdC8KA6AgXhGsqQAvZmvKpwOAE6YrW-0Vru-tGa3TZWYThSPQ4M_mN6sODataUkXVTBN4eBGL4sbMpq9GnecMw2bBLSrSC44bzAr5ZwB5KY35yoejpGVaXVGIsSmdtoVb_ocoxdvQ6TNb5Ej8quFgKdAwpRev-9k6wml2liqvU7ColZ_r143H_sQcbsd9CMMp0</recordid><startdate>20090101</startdate><enddate>20090101</enddate><creator>Sasaki, Katsunori</creator><creator>Ichikawa, Hinako</creator><creator>Takei, Shunsuke</creator><creator>No, Hee Sung</creator><creator>Tomotsune, Daihachiro</creator><creator>Kano, Yoshiya</creator><creator>Yokoyama, Tadayuki</creator><creator>Sirasawa, Sakiko</creator><creator>Mogi, Akimi</creator><creator>Yoshie, Susumu</creator><creator>Sasaki, Shujiro</creator><creator>Yamada, Satoshi</creator><creator>Matsumoto, Ken</creator><creator>Mizuguchi, Masahiro</creator><creator>Yue, Fengming</creator><creator>Tanaka, Yoshiki</creator><general>John Wiley & Sons, Inc</general><general>TheScientificWorldJOURNAL</general><general>Hindawi Limited</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20090101</creationdate><title>Hepatocyte differentiation from human ES cells using the simple embryoid body formation method and the staged-additional cocktail</title><author>Sasaki, Katsunori ; Ichikawa, Hinako ; Takei, Shunsuke ; No, Hee Sung ; Tomotsune, Daihachiro ; Kano, Yoshiya ; Yokoyama, Tadayuki ; Sirasawa, Sakiko ; Mogi, Akimi ; Yoshie, Susumu ; Sasaki, Shujiro ; Yamada, Satoshi ; Matsumoto, Ken ; Mizuguchi, Masahiro ; Yue, Fengming ; Tanaka, Yoshiki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c622t-5b72b342d6ad3430586c0625a20d95f94ad96f9620431fb30fc4ae0fb9e1f8f33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Base Sequence</topic><topic>Cell differentiation</topic><topic>Cell Line</topic><topic>DNA Primers</topic><topic>Embryonic development</topic><topic>Embryonic stem cells</topic><topic>Embryonic Stem Cells - cytology</topic><topic>Embryonic Stem Cells - ultrastructure</topic><topic>Health aspects</topic><topic>Hepatocytes - cytology</topic><topic>Humans</topic><topic>Liver cells</topic><topic>Microscopy, Electron, Transmission</topic><topic>Physiological aspects</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Short Communication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sasaki, Katsunori</creatorcontrib><creatorcontrib>Ichikawa, Hinako</creatorcontrib><creatorcontrib>Takei, Shunsuke</creatorcontrib><creatorcontrib>No, Hee Sung</creatorcontrib><creatorcontrib>Tomotsune, Daihachiro</creatorcontrib><creatorcontrib>Kano, Yoshiya</creatorcontrib><creatorcontrib>Yokoyama, Tadayuki</creatorcontrib><creatorcontrib>Sirasawa, Sakiko</creatorcontrib><creatorcontrib>Mogi, Akimi</creatorcontrib><creatorcontrib>Yoshie, Susumu</creatorcontrib><creatorcontrib>Sasaki, Shujiro</creatorcontrib><creatorcontrib>Yamada, Satoshi</creatorcontrib><creatorcontrib>Matsumoto, Ken</creatorcontrib><creatorcontrib>Mizuguchi, Masahiro</creatorcontrib><creatorcontrib>Yue, Fengming</creatorcontrib><creatorcontrib>Tanaka, Yoshiki</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>TheScientificWorld</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sasaki, Katsunori</au><au>Ichikawa, Hinako</au><au>Takei, Shunsuke</au><au>No, Hee Sung</au><au>Tomotsune, Daihachiro</au><au>Kano, Yoshiya</au><au>Yokoyama, Tadayuki</au><au>Sirasawa, Sakiko</au><au>Mogi, Akimi</au><au>Yoshie, Susumu</au><au>Sasaki, Shujiro</au><au>Yamada, Satoshi</au><au>Matsumoto, Ken</au><au>Mizuguchi, Masahiro</au><au>Yue, Fengming</au><au>Tanaka, Yoshiki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Hepatocyte differentiation from human ES cells using the simple embryoid body formation method and the staged-additional cocktail</atitle><jtitle>TheScientificWorld</jtitle><addtitle>ScientificWorldJournal</addtitle><date>2009-01-01</date><risdate>2009</risdate><volume>9</volume><spage>884</spage><epage>890</epage><pages>884-890</pages><issn>1537-744X</issn><issn>2356-6140</issn><eissn>1537-744X</eissn><abstract>To induce hepatocytes from human embryonic stem (hES) cells easily and effectively, a simple suspension culture method that separates ES colonies with a scraper and transfers them into newly developed, nonadherent MPC (2-methacryloyloxyethyl phosphorylcholine) plates, and the staged-additional cocktail method, including growth factors, cytokines, and Lanford serum-free medium, were developed and evaluated mainly by morphological analysis. 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| source | Open Access: PubMed Central; Wiley-Blackwell Titles (Open access) |
| subjects | Base Sequence Cell differentiation Cell Line DNA Primers Embryonic development Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - ultrastructure Health aspects Hepatocytes - cytology Humans Liver cells Microscopy, Electron, Transmission Physiological aspects Reverse Transcriptase Polymerase Chain Reaction Short Communication |
| title | Hepatocyte differentiation from human ES cells using the simple embryoid body formation method and the staged-additional cocktail |
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