Mir‐122 upregulation and let‐7f downregulation combination: The effects on hepatic differentiation of hiPSCs on the PCL‐Gel‐HA nanofibrous scaffold
Cell therapy and tissue engineering as promising candidates for the liver transplantation dilemma are of special interest. Induced pluripotent stem cells (iPSCs) are one of the best sources in this field, but their differentiation methods to hepatocytes have remained challenging. We transduced human...
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| Veröffentlicht in: | Journal of cellular and molecular medicine 2022-10, Vol.26 (20), p.5235-5245 |
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| creator | Parvanak, Maliheh Mostafavi‐Pour, Zohreh Soleimani, Masoud Atashi, Amir Arefian, Ehsan Esmaeili, Elaheh |
| description | Cell therapy and tissue engineering as promising candidates for the liver transplantation dilemma are of special interest. Induced pluripotent stem cells (iPSCs) are one of the best sources in this field, but their differentiation methods to hepatocytes have remained challenging. We transduced human iPSCs (hiPSCs) with miR‐122 and off‐let‐7f (hiPSCsmiR‐122 + off‐let‐7f) to evaluate how they can differentiate hiPSCs to hepatocyte‐like cells (HLCs) without any extrinsic growth factor. Additionally, we studied the effect of Poly ɛ‐caprolactone‐gelatin‐hyaluronic acid (PCL‐Gel‐HA) nanofibrous scaffold as an extracellular matrix (ECM) simulator on differentiation improvement. Definitive endoderm markers (FOXA2 and SOX17), as well as hepatic markers (AFP, Albumin, CK18, HNF4α) expression, were significantly higher in hiPSCsmiR‐122 + off‐let‐7f derived HLCs (hiPSCs‐HLCs) compared to the control group (miR‐scramble transduced hiPSCs: hiPSCsscramble). hiPSCs‐HLCs indicated hepatocyte morphological characteristics and positive immunostaining for AFP, Albumin and HNF4α. Albumin and urea secretion were significantly higher in hiPSCs‐HLCs than hiPSCsscramble. Comparing these markers in the PCL‐Gel‐HA group with the tissue culture plate (TCP) group revealed that PCL‐Gel‐HA could improve differentiation towards HLCs significantly. Regarding our results, these microRNAs can be used to differentiate hiPSCs to the functional hepatocytes for disease modelling, drug screening and cell‐based therapy in future studies. |
| doi_str_mv | 10.1111/jcmm.17552 |
| format | Article |
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Induced pluripotent stem cells (iPSCs) are one of the best sources in this field, but their differentiation methods to hepatocytes have remained challenging. We transduced human iPSCs (hiPSCs) with miR‐122 and off‐let‐7f (hiPSCsmiR‐122 + off‐let‐7f) to evaluate how they can differentiate hiPSCs to hepatocyte‐like cells (HLCs) without any extrinsic growth factor. Additionally, we studied the effect of Poly ɛ‐caprolactone‐gelatin‐hyaluronic acid (PCL‐Gel‐HA) nanofibrous scaffold as an extracellular matrix (ECM) simulator on differentiation improvement. Definitive endoderm markers (FOXA2 and SOX17), as well as hepatic markers (AFP, Albumin, CK18, HNF4α) expression, were significantly higher in hiPSCsmiR‐122 + off‐let‐7f derived HLCs (hiPSCs‐HLCs) compared to the control group (miR‐scramble transduced hiPSCs: hiPSCsscramble). hiPSCs‐HLCs indicated hepatocyte morphological characteristics and positive immunostaining for AFP, Albumin and HNF4α. Albumin and urea secretion were significantly higher in hiPSCs‐HLCs than hiPSCsscramble. Comparing these markers in the PCL‐Gel‐HA group with the tissue culture plate (TCP) group revealed that PCL‐Gel‐HA could improve differentiation towards HLCs significantly. Regarding our results, these microRNAs can be used to differentiate hiPSCs to the functional hepatocytes for disease modelling, drug screening and cell‐based therapy in future studies.</description><identifier>ISSN: 1582-1838</identifier><identifier>EISSN: 1582-4934</identifier><identifier>DOI: 10.1111/jcmm.17552</identifier><identifier>PMID: 36098216</identifier><language>eng</language><publisher>Chichester: John Wiley & Sons, Inc</publisher><subject>Albumin ; Cell adhesion & migration ; Cell culture ; Cell differentiation ; Cell growth ; Cell therapy ; Contact angle ; Drug screening ; Endoderm ; Extracellular matrix ; Fibroblasts ; Gelatin ; Gene expression ; Growth factors ; hepatic differentiation ; Hepatocytes ; Hyaluronic acid ; induced pluripotent stem cells ; let‐7f ; Liver ; Liver transplantation ; miRNA ; miR‐122 ; Morphology ; nanofibrous scaffold ; Original ; Physical characteristics ; Pluripotency ; Scanning electron microscopy ; Signal transduction ; Stem cells ; Tissue culture ; Tissue engineering</subject><ispartof>Journal of cellular and molecular medicine, 2022-10, Vol.26 (20), p.5235-5245</ispartof><rights>2022 The Authors. published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.</rights><rights>2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c3142-a9bede09129768bccc19bab11497f75c07b1504366c8bddafa7b3080f0d8b7f73</cites><orcidid>0000-0003-0324-9808 ; 0000-0003-4820-8559 ; 0000-0002-3779-177X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9575133/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9575133/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,1417,11562,27924,27925,45574,45575,46052,46476,53791,53793</link.rule.ids></links><search><creatorcontrib>Parvanak, Maliheh</creatorcontrib><creatorcontrib>Mostafavi‐Pour, Zohreh</creatorcontrib><creatorcontrib>Soleimani, Masoud</creatorcontrib><creatorcontrib>Atashi, Amir</creatorcontrib><creatorcontrib>Arefian, Ehsan</creatorcontrib><creatorcontrib>Esmaeili, Elaheh</creatorcontrib><title>Mir‐122 upregulation and let‐7f downregulation combination: The effects on hepatic differentiation of hiPSCs on the PCL‐Gel‐HA nanofibrous scaffold</title><title>Journal of cellular and molecular medicine</title><description>Cell therapy and tissue engineering as promising candidates for the liver transplantation dilemma are of special interest. Induced pluripotent stem cells (iPSCs) are one of the best sources in this field, but their differentiation methods to hepatocytes have remained challenging. We transduced human iPSCs (hiPSCs) with miR‐122 and off‐let‐7f (hiPSCsmiR‐122 + off‐let‐7f) to evaluate how they can differentiate hiPSCs to hepatocyte‐like cells (HLCs) without any extrinsic growth factor. Additionally, we studied the effect of Poly ɛ‐caprolactone‐gelatin‐hyaluronic acid (PCL‐Gel‐HA) nanofibrous scaffold as an extracellular matrix (ECM) simulator on differentiation improvement. Definitive endoderm markers (FOXA2 and SOX17), as well as hepatic markers (AFP, Albumin, CK18, HNF4α) expression, were significantly higher in hiPSCsmiR‐122 + off‐let‐7f derived HLCs (hiPSCs‐HLCs) compared to the control group (miR‐scramble transduced hiPSCs: hiPSCsscramble). hiPSCs‐HLCs indicated hepatocyte morphological characteristics and positive immunostaining for AFP, Albumin and HNF4α. Albumin and urea secretion were significantly higher in hiPSCs‐HLCs than hiPSCsscramble. Comparing these markers in the PCL‐Gel‐HA group with the tissue culture plate (TCP) group revealed that PCL‐Gel‐HA could improve differentiation towards HLCs significantly. Regarding our results, these microRNAs can be used to differentiate hiPSCs to the functional hepatocytes for disease modelling, drug screening and cell‐based therapy in future studies.</description><subject>Albumin</subject><subject>Cell adhesion & migration</subject><subject>Cell culture</subject><subject>Cell differentiation</subject><subject>Cell growth</subject><subject>Cell therapy</subject><subject>Contact angle</subject><subject>Drug screening</subject><subject>Endoderm</subject><subject>Extracellular matrix</subject><subject>Fibroblasts</subject><subject>Gelatin</subject><subject>Gene expression</subject><subject>Growth factors</subject><subject>hepatic differentiation</subject><subject>Hepatocytes</subject><subject>Hyaluronic acid</subject><subject>induced pluripotent stem cells</subject><subject>let‐7f</subject><subject>Liver</subject><subject>Liver transplantation</subject><subject>miRNA</subject><subject>miR‐122</subject><subject>Morphology</subject><subject>nanofibrous scaffold</subject><subject>Original</subject><subject>Physical characteristics</subject><subject>Pluripotency</subject><subject>Scanning electron microscopy</subject><subject>Signal transduction</subject><subject>Stem cells</subject><subject>Tissue culture</subject><subject>Tissue 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upregulation and let‐7f downregulation combination: The effects on hepatic differentiation of hiPSCs on the PCL‐Gel‐HA nanofibrous scaffold</title><author>Parvanak, Maliheh ; Mostafavi‐Pour, Zohreh ; Soleimani, Masoud ; Atashi, Amir ; Arefian, Ehsan ; Esmaeili, Elaheh</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3142-a9bede09129768bccc19bab11497f75c07b1504366c8bddafa7b3080f0d8b7f73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Albumin</topic><topic>Cell adhesion & migration</topic><topic>Cell culture</topic><topic>Cell differentiation</topic><topic>Cell growth</topic><topic>Cell therapy</topic><topic>Contact angle</topic><topic>Drug screening</topic><topic>Endoderm</topic><topic>Extracellular matrix</topic><topic>Fibroblasts</topic><topic>Gelatin</topic><topic>Gene expression</topic><topic>Growth factors</topic><topic>hepatic differentiation</topic><topic>Hepatocytes</topic><topic>Hyaluronic acid</topic><topic>induced pluripotent stem cells</topic><topic>let‐7f</topic><topic>Liver</topic><topic>Liver transplantation</topic><topic>miRNA</topic><topic>miR‐122</topic><topic>Morphology</topic><topic>nanofibrous scaffold</topic><topic>Original</topic><topic>Physical characteristics</topic><topic>Pluripotency</topic><topic>Scanning electron microscopy</topic><topic>Signal transduction</topic><topic>Stem cells</topic><topic>Tissue culture</topic><topic>Tissue engineering</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Parvanak, Maliheh</creatorcontrib><creatorcontrib>Mostafavi‐Pour, Zohreh</creatorcontrib><creatorcontrib>Soleimani, Masoud</creatorcontrib><creatorcontrib>Atashi, Amir</creatorcontrib><creatorcontrib>Arefian, Ehsan</creatorcontrib><creatorcontrib>Esmaeili, Elaheh</creatorcontrib><collection>Wiley Online Library Open Access</collection><collection>Wiley 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differentiation of hiPSCs on the PCL‐Gel‐HA nanofibrous scaffold</atitle><jtitle>Journal of cellular and molecular medicine</jtitle><date>2022-10</date><risdate>2022</risdate><volume>26</volume><issue>20</issue><spage>5235</spage><epage>5245</epage><pages>5235-5245</pages><issn>1582-1838</issn><eissn>1582-4934</eissn><abstract>Cell therapy and tissue engineering as promising candidates for the liver transplantation dilemma are of special interest. Induced pluripotent stem cells (iPSCs) are one of the best sources in this field, but their differentiation methods to hepatocytes have remained challenging. We transduced human iPSCs (hiPSCs) with miR‐122 and off‐let‐7f (hiPSCsmiR‐122 + off‐let‐7f) to evaluate how they can differentiate hiPSCs to hepatocyte‐like cells (HLCs) without any extrinsic growth factor. Additionally, we studied the effect of Poly ɛ‐caprolactone‐gelatin‐hyaluronic acid (PCL‐Gel‐HA) nanofibrous scaffold as an extracellular matrix (ECM) simulator on differentiation improvement. Definitive endoderm markers (FOXA2 and SOX17), as well as hepatic markers (AFP, Albumin, CK18, HNF4α) expression, were significantly higher in hiPSCsmiR‐122 + off‐let‐7f derived HLCs (hiPSCs‐HLCs) compared to the control group (miR‐scramble transduced hiPSCs: hiPSCsscramble). hiPSCs‐HLCs indicated hepatocyte morphological characteristics and positive immunostaining for AFP, Albumin and HNF4α. Albumin and urea secretion were significantly higher in hiPSCs‐HLCs than hiPSCsscramble. Comparing these markers in the PCL‐Gel‐HA group with the tissue culture plate (TCP) group revealed that PCL‐Gel‐HA could improve differentiation towards HLCs significantly. Regarding our results, these microRNAs can be used to differentiate hiPSCs to the functional hepatocytes for disease modelling, drug screening and cell‐based therapy in future studies.</abstract><cop>Chichester</cop><pub>John Wiley & Sons, Inc</pub><pmid>36098216</pmid><doi>10.1111/jcmm.17552</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0003-0324-9808</orcidid><orcidid>https://orcid.org/0000-0003-4820-8559</orcidid><orcidid>https://orcid.org/0000-0002-3779-177X</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Albumin Cell adhesion & migration Cell culture Cell differentiation Cell growth Cell therapy Contact angle Drug screening Endoderm Extracellular matrix Fibroblasts Gelatin Gene expression Growth factors hepatic differentiation Hepatocytes Hyaluronic acid induced pluripotent stem cells let‐7f Liver Liver transplantation miRNA miR‐122 Morphology nanofibrous scaffold Original Physical characteristics Pluripotency Scanning electron microscopy Signal transduction Stem cells Tissue culture Tissue engineering |
| title | Mir‐122 upregulation and let‐7f downregulation combination: The effects on hepatic differentiation of hiPSCs on the PCL‐Gel‐HA nanofibrous scaffold |
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