Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor
Identifieur interne : 000339 ( Pmc/Curation ); précédent : 000338; suivant : 000340Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor
Auteurs : Nora Freyer ; Fanny Knöspel ; Nadja Strahl ; Leila Amini ; Petra Schrade ; Sebastian Bachmann ; Georg Damm ; Daniel Seehofer ; Frank Jacobs ; Mario Monshouwer ; Katrin ZeilingerSource :
- BioResearch Open Access [ 2164-7844 ] ; 2016.
Abstract
The hepatic differentiation of human induced pluripotent stem cells (hiPSC) holds great potential for application in regenerative medicine, pharmacological drug screening, and toxicity testing. However, full maturation of hiPSC into functional hepatocytes has not yet been achieved. In this study, we investigated the potential of a dynamic three-dimensional (3D) hollow fiber membrane bioreactor technology to improve the hepatic differentiation of hiPSC in comparison to static two-dimensional (2D) cultures. A total of 100 × 106 hiPSC were seeded into each 3D bioreactor (
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DOI: 10.1089/biores.2016.0027
PubMed: 27610270
PubMed Central: 5003005
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor</title><author><name sortKey="Freyer, Nora" sort="Freyer, Nora" uniqKey="Freyer N" first="Nora" last="Freyer">Nora Freyer</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Knospel, Fanny" sort="Knospel, Fanny" uniqKey="Knospel F" first="Fanny" last="Knöspel">Fanny Knöspel</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Strahl, Nadja" sort="Strahl, Nadja" uniqKey="Strahl N" first="Nadja" last="Strahl">Nadja Strahl</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Amini, Leila" sort="Amini, Leila" uniqKey="Amini L" first="Leila" last="Amini">Leila Amini</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Schrade, Petra" sort="Schrade, Petra" uniqKey="Schrade P" first="Petra" last="Schrade">Petra Schrade</name><affiliation><nlm:aff id="aff2"></nlm:aff></affiliation></author><author><name sortKey="Bachmann, Sebastian" sort="Bachmann, Sebastian" uniqKey="Bachmann S" first="Sebastian" last="Bachmann">Sebastian Bachmann</name><affiliation><nlm:aff id="aff2"></nlm:aff></affiliation></author><author><name sortKey="Damm, Georg" sort="Damm, Georg" uniqKey="Damm G" first="Georg" last="Damm">Georg Damm</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff id="aff4"></nlm:aff></affiliation></author><author><name sortKey="Seehofer, Daniel" sort="Seehofer, Daniel" uniqKey="Seehofer D" first="Daniel" last="Seehofer">Daniel Seehofer</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff id="aff4"></nlm:aff></affiliation></author><author><name sortKey="Jacobs, Frank" sort="Jacobs, Frank" uniqKey="Jacobs F" first="Frank" last="Jacobs">Frank Jacobs</name><affiliation><nlm:aff id="aff5"></nlm:aff></affiliation></author><author><name sortKey="Monshouwer, Mario" sort="Monshouwer, Mario" uniqKey="Monshouwer M" first="Mario" last="Monshouwer">Mario Monshouwer</name><affiliation><nlm:aff id="aff5"></nlm:aff></affiliation></author><author><name sortKey="Zeilinger, Katrin" sort="Zeilinger, Katrin" uniqKey="Zeilinger K" first="Katrin" last="Zeilinger">Katrin Zeilinger</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">27610270</idno><idno type="pmc">5003005</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003005</idno><idno type="RBID">PMC:5003005</idno><idno type="doi">10.1089/biores.2016.0027</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000339</idno><idno type="wicri:Area/Pmc/Curation">000339</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor</title><author><name sortKey="Freyer, Nora" sort="Freyer, Nora" uniqKey="Freyer N" first="Nora" last="Freyer">Nora Freyer</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Knospel, Fanny" sort="Knospel, Fanny" uniqKey="Knospel F" first="Fanny" last="Knöspel">Fanny Knöspel</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Strahl, Nadja" sort="Strahl, Nadja" uniqKey="Strahl N" first="Nadja" last="Strahl">Nadja Strahl</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Amini, Leila" sort="Amini, Leila" uniqKey="Amini L" first="Leila" last="Amini">Leila Amini</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Schrade, Petra" sort="Schrade, Petra" uniqKey="Schrade P" first="Petra" last="Schrade">Petra Schrade</name><affiliation><nlm:aff id="aff2"></nlm:aff></affiliation></author><author><name sortKey="Bachmann, Sebastian" sort="Bachmann, Sebastian" uniqKey="Bachmann S" first="Sebastian" last="Bachmann">Sebastian Bachmann</name><affiliation><nlm:aff id="aff2"></nlm:aff></affiliation></author><author><name sortKey="Damm, Georg" sort="Damm, Georg" uniqKey="Damm G" first="Georg" last="Damm">Georg Damm</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff id="aff4"></nlm:aff></affiliation></author><author><name sortKey="Seehofer, Daniel" sort="Seehofer, Daniel" uniqKey="Seehofer D" first="Daniel" last="Seehofer">Daniel Seehofer</name><affiliation><nlm:aff id="aff3"></nlm:aff></affiliation><affiliation><nlm:aff id="aff4"></nlm:aff></affiliation></author><author><name sortKey="Jacobs, Frank" sort="Jacobs, Frank" uniqKey="Jacobs F" first="Frank" last="Jacobs">Frank Jacobs</name><affiliation><nlm:aff id="aff5"></nlm:aff></affiliation></author><author><name sortKey="Monshouwer, Mario" sort="Monshouwer, Mario" uniqKey="Monshouwer M" first="Mario" last="Monshouwer">Mario Monshouwer</name><affiliation><nlm:aff id="aff5"></nlm:aff></affiliation></author><author><name sortKey="Zeilinger, Katrin" sort="Zeilinger, Katrin" uniqKey="Zeilinger K" first="Katrin" last="Zeilinger">Katrin Zeilinger</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">BioResearch Open Access</title><idno type="ISSN">2164-7844</idno><idno type="eISSN">2164-7860</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Abstract</title><p>The hepatic differentiation of human induced pluripotent stem cells (hiPSC) holds great potential for application in regenerative medicine, pharmacological drug screening, and toxicity testing. However, full maturation of hiPSC into functional hepatocytes has not yet been achieved. In this study, we investigated the potential of a dynamic three-dimensional (3D) hollow fiber membrane bioreactor technology to improve the hepatic differentiation of hiPSC in comparison to static two-dimensional (2D) cultures. A total of 100 × 10<sup>6</sup> hiPSC were seeded into each 3D bioreactor (<italic>n</italic> = 3). Differentiation into definitive endoderm (DE) was induced by adding activin A, Wnt3a, and sodium butyrate to the culture medium. For further maturation, hepatocyte growth factor and oncostatin M were added. The same differentiation protocol was applied to hiPSC maintained in 2D cultures. Secretion of alpha-fetoprotein (AFP), a marker for DE, was significantly (<italic>p</italic> < 0.05) higher in 2D cultures, while secretion of albumin, a typical characteristic for mature hepatocytes, was higher after hepatic differentiation of hiPSC in 3D bioreactors. Functional analysis of multiple cytochrome P450 (CYP) isoenzymes showed activity of CYP1A2, CYP2B6, and CYP3A4 in both groups, although at a lower level compared to primary human hepatocytes (PHH). CYP2B6 activities were significantly (<italic>p</italic> < 0.05) higher in 3D bioreactors compared with 2D cultures, which is in line with results from gene expression. Immunofluorescence staining showed that the majority of cells was positive for albumin, cytokeratin 18 (CK18), and hepatocyte nuclear factor 4-alpha (HNF4A) at the end of the differentiation process. In addition, cytokeratin 19 (CK19) staining revealed the formation of bile duct-like structures in 3D bioreactors similar to native liver tissue. The results indicate a better maturation of hiPSC in the 3D bioreactor system compared to 2D cultures and emphasize the potential of dynamic 3D culture systems in stem cell differentiation approaches for improved formation of differentiated tissue structures.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Kola, I" uniqKey="Kola I">I Kola</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lewis, Df" uniqKey="Lewis D">DF Lewis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Giri, S" uniqKey="Giri S">S Giri</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ingelman Sundberg, M" uniqKey="Ingelman Sundberg M">M Ingelman-Sundberg</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Takayama, K" uniqKey="Takayama K">K Takayama</name></author></analytic></biblStruct><biblStruct><analytic><author><name 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Biores Open Access</journal-id><journal-id journal-id-type="iso-abbrev">Biores Open Access</journal-id><journal-id journal-id-type="publisher-id">biores</journal-id><journal-title-group><journal-title>BioResearch Open Access</journal-title></journal-title-group><issn pub-type="ppub">2164-7844</issn><issn pub-type="epub">2164-7860</issn><publisher><publisher-name>Mary Ann Liebert, Inc.</publisher-name><publisher-loc>140 Huguenot Street, 3rd FloorNew Rochelle, NY 10801USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">27610270</article-id><article-id pub-id-type="pmc">5003005</article-id><article-id pub-id-type="publisher-id">10.1089/biores.2016.0027</article-id><article-id pub-id-type="doi">10.1089/biores.2016.0027</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Research Article</subject></subj-group></article-categories><title-group><article-title>Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Freyer</surname><given-names>Nora</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Knöspel</surname><given-names>Fanny</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Strahl</surname><given-names>Nadja</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Amini</surname><given-names>Leila</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Schrade</surname><given-names>Petra</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Bachmann</surname><given-names>Sebastian</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Damm</surname><given-names>Georg</given-names></name><xref ref-type="aff" rid="aff3"><sup>3,</sup></xref><xref ref-type="aff" rid="aff4"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Seehofer</surname><given-names>Daniel</given-names></name><xref ref-type="aff" rid="aff3"><sup>3,</sup></xref><xref ref-type="aff" rid="aff4"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Jacobs</surname><given-names>Frank</given-names></name><xref ref-type="aff" rid="aff5"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Monshouwer</surname><given-names>Mario</given-names></name><xref ref-type="aff" rid="aff5"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Zeilinger</surname><given-names>Katrin</given-names></name><xref ref-type="aff" rid="aff1"><sup>1,</sup></xref><xref ref-type="corresp" rid="corr1"><sup>*</sup></xref></contrib><aff id="aff1"><label><sup>1</sup></label>Bioreactor Group, Berlin Brandenburg Center for Regenerative Therapies (BCRT),<institution>Charité—Universitätsmedizin Berlin</institution>, Berlin,<country>Germany</country>.</aff><aff id="aff2"><label><sup>2</sup></label>Charité Centrum Grundlagenmedizin, Institut für Vegetative Anatomie,<institution>Charité—Universitätsmedizin Berlin</institution>, Berlin,<country>Germany</country>.</aff><aff id="aff3"><label><sup>3</sup></label>Department of General-, Visceral- and Transplantation Surgery,<institution>Charité—Universitätsmedizin Berlin</institution>, Berlin,<country>Germany</country>.</aff><aff id="aff4"><label><sup>4</sup></label>Department of Hepatobiliary Surgery and Visceral Transplantation,<institution>University of Leipzig</institution>, Leipzig,<country>Germany</country>.</aff><aff id="aff5"><label><sup>5</sup></label><institution>Janssen Research and Development</institution>, Beerse,<country>Belgium</country>.</aff></contrib-group><author-notes><fn id="fn1" fn-type="presented-at"><p>Part of this work was previously presented at the following meeting: Freyer N, Strahl N, Knöspel F, Urbaniak T, Zeilinger K. Hepatic differentiation of hiPSC in a 3D multicompartment bioreactor, 30th Annual Meeting of the German Association of the Study of the Liver (GASL), 2014, 24 and 25th January, Tübingen, Germany, abstract published in Gastroenterol 52, p. 3.12.</p></fn><corresp id="corr1"><label><sup>*</sup></label>Address correspondence to: Dr. med. vet. Katrin Zeilinger, Bioreactor Group, Berlin Brandenburg Center for Regenerative Therapies (BCRT), Charité—Universitätsmedizin Berlin, Campus Virchow-Klinikum, Augustenburger Platz 1, Berlin 13353, Germany, E-mail: <email xlink:href="mailto:katrin.zeilinger@charite.de">katrin.zeilinger@charite.de</email></corresp></author-notes><pub-date pub-type="epub"><day>01</day><month>8</month><year>2016</year><pmc-comment>string-date: August 2016</pmc-comment>
</pub-date><pub-date pub-type="collection"><year>2016</year></pub-date><pub-date pub-type="pmc-release"><day>01</day><month>8</month><year>2016</year><pmc-comment>string-date: August 2016</pmc-comment>
</pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>5</volume><issue>1</issue><fpage>235</fpage><lpage>248</lpage><permissions><copyright-statement>© Nora Freyer <italic>et al</italic>. 2016; Published by Mary Ann Liebert, Inc.</copyright-statement><copyright-year>2016</copyright-year><license license-type="open-access"><license-p>This Open Access article is distributed under the terms of the Creative Commons License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0">http://creativecommons.org/licenses/by/4.0</ext-link>), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.</license-p></license></permissions><self-uri content-type="pdf" xlink:type="simple" xlink:href="biores.2016.0027.pdf"></self-uri><abstract><title>Abstract</title><p>The hepatic differentiation of human induced pluripotent stem cells (hiPSC) holds great potential for application in regenerative medicine, pharmacological drug screening, and toxicity testing. However, full maturation of hiPSC into functional hepatocytes has not yet been achieved. In this study, we investigated the potential of a dynamic three-dimensional (3D) hollow fiber membrane bioreactor technology to improve the hepatic differentiation of hiPSC in comparison to static two-dimensional (2D) cultures. A total of 100 × 10<sup>6</sup> hiPSC were seeded into each 3D bioreactor (<italic>n</italic> = 3). Differentiation into definitive endoderm (DE) was induced by adding activin A, Wnt3a, and sodium butyrate to the culture medium. For further maturation, hepatocyte growth factor and oncostatin M were added. The same differentiation protocol was applied to hiPSC maintained in 2D cultures. Secretion of alpha-fetoprotein (AFP), a marker for DE, was significantly (<italic>p</italic> < 0.05) higher in 2D cultures, while secretion of albumin, a typical characteristic for mature hepatocytes, was higher after hepatic differentiation of hiPSC in 3D bioreactors. Functional analysis of multiple cytochrome P450 (CYP) isoenzymes showed activity of CYP1A2, CYP2B6, and CYP3A4 in both groups, although at a lower level compared to primary human hepatocytes (PHH). CYP2B6 activities were significantly (<italic>p</italic> < 0.05) higher in 3D bioreactors compared with 2D cultures, which is in line with results from gene expression. Immunofluorescence staining showed that the majority of cells was positive for albumin, cytokeratin 18 (CK18), and hepatocyte nuclear factor 4-alpha (HNF4A) at the end of the differentiation process. In addition, cytokeratin 19 (CK19) staining revealed the formation of bile duct-like structures in 3D bioreactors similar to native liver tissue. The results indicate a better maturation of hiPSC in the 3D bioreactor system compared to 2D cultures and emphasize the potential of dynamic 3D culture systems in stem cell differentiation approaches for improved formation of differentiated tissue structures.</p></abstract><kwd-group kwd-group-type="author"><title><bold>Keywords:</bold> </title><kwd>stem cells</kwd><kwd>tissue engineering</kwd></kwd-group><counts><fig-count count="5"></fig-count><table-count count="3"></table-count><ref-count count="59"></ref-count><page-count count="14"></page-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Curation/RBID.i -Sk "pubmed:27610270" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd \
| NlmPubMed2Wicri -a OpenAccessBelV2
| This area was generated with Dilib version V0.6.25. |  |