MafA Overexpression: A New Efficient Protocol for In Vitro Differentiation of Adipose-Derived Mesenchymal Stem Cells into Functional Insulin-Producing Cells
We proposed a novel differentiation method for the efficient differentiation of adipose-derived mesenchymal stem cells (ADMSCs) into functional insulin-producing cells (IPCs) based on overexpression. In this experimental study, a eukaryotic expression vector containing [ /pcDNA3.1(+)] was constructe...
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| Veröffentlicht in: | Cell journal (Yakhteh) 2019-07, Vol.21 (2), p.169-178 |
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| creator | Dayer, Dian Tabandeh, Mohammad Reza Moghimipour, Eskandar Hashemi Tabar, Mahmood Ghadiri, AtaA Allah Bakhshi, Elham Orazizadeh, Mahmoud Ghafari, Mohammad Ali |
| description | We proposed a novel differentiation method for the efficient differentiation of adipose-derived mesenchymal stem cells (ADMSCs) into functional insulin-producing cells (IPCs) based on
overexpression.
In this experimental study, a eukaryotic expression vector containing
[
/pcDNA3.1(+)] was constructed and purified. ADMSCs were differentiated into IPCs. ADMSCs were assigned in two groups including control (C), and the
overexpressed (
+) groups. The ADMSCs were transfected by
/pcDNA 3.1(+) at day 10 of the differentiation. Differentiated cells were analyzed for the expression of multiple β cell specific genes (
,
,
,
,
,
, and
) using real-time polymerase chain reaction (PCR). The insulin secretion potency of the differentiated cells in response to glucose exposure was also determined using an enzyme-linked immunosorbent assay (ELISA) method and Dithizone (DTZ) staining. The IPCs from the control manipulated group, and un-differentiated ADMSCs group were transplanted to streptozotocin (STZ)-diabetic rats. Rats were monitored for blood glucose and insulin concentration.
The results revealed that ADMSCs were successfully differentiated into IPCs through the 14 day differentiation protocol. The expression of β-cell specific genes in
+ IPCs was higher than in control cells. Glucose-induced insulin secretion after the exposure of IPCs to glucose was higher in
+ group than the control group. The STZdiabetic rats showed an ability to secrete insulin and apparent hyperglycemic condition adjustment after transplantation of the control IPCs. The mean insulin concentration of diabetic rats that were transplanted by manipulated IPCs was significantly higher than ADMSCs-transplanted rats; however, no effect was observed in the concentration of bloodn glucose.
The overexpression of
can be used as a novel promising approach for the efficient production of IPCs from ADMSCs in vitro. However, the future therapeutic use of the
+ IPCs in diabetic animals needs further investigations. |
| doi_str_mv | 10.22074/cellj.2019.5669 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6397604</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2187525391</sourcerecordid><originalsourceid>FETCH-LOGICAL-p294t-ac35b009ad3fc4fcb8bdaa95087bda77423b14570c5d0671b4c30f534d8218d53</originalsourceid><addsrcrecordid>eNpdkUtv1DAUhS0EolXpnhWyxIZNBsfPmAXSaNrCSC1F4rGNHD9ajxI7tZ2B_hd-LK5aKsAbX-l-59xjXwBetmiFMRL0rbbjuFth1MoV41w-AYcY465hXUufPtaIH4DjnHeoHo5wK_FzcEBQhxmW6BD8ulBuDS_3Ntmfc7I5-xjewTX8ZH_AU-e89jYU-DnFEnUcoYsJbgP87kuK8MQ7V3WheFWqDEYH18bPMdvmxCa_twZe2GyDvr6d1Ai_FDvBTc2coQ8lwrMl6DtdbW1DXkYfmjrHLNqHq3vuBXjm1Jjt8cN9BL6dnX7dfGzOLz9sN-vzZsaSlkZpwgaEpDLEaer00A1GKclQJ2ohBMVkaCkTSDODuGgHqglyjFDT4bYzjByB9_e-8zJM1uj6pKTGfk5-Uum2j8r3_3aCv-6v4r7nRAqOaDV482CQ4s1ic-knn-_Wo4KNS-7rGMEwI7Kt6Ov_0F1cUv2ESnUUSc4kF5V69Xeixyh_Fkd-A-Dunu4</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2840965967</pqid></control><display><type>article</type><title>MafA Overexpression: A New Efficient Protocol for In Vitro Differentiation of Adipose-Derived Mesenchymal Stem Cells into Functional Insulin-Producing Cells</title><source>DOAJ Directory of Open Access Journals</source><source>PubMed Central Open Access</source><source>PubMed Central</source><creator>Dayer, Dian ; Tabandeh, Mohammad Reza ; Moghimipour, Eskandar ; Hashemi Tabar, Mahmood ; Ghadiri, AtaA ; Allah Bakhshi, Elham ; Orazizadeh, Mahmoud ; Ghafari, Mohammad Ali</creator><creatorcontrib>Dayer, Dian ; Tabandeh, Mohammad Reza ; Moghimipour, Eskandar ; Hashemi Tabar, Mahmood ; Ghadiri, AtaA ; Allah Bakhshi, Elham ; Orazizadeh, Mahmoud ; Ghafari, Mohammad Ali</creatorcontrib><description>We proposed a novel differentiation method for the efficient differentiation of adipose-derived mesenchymal stem cells (ADMSCs) into functional insulin-producing cells (IPCs) based on
overexpression.
In this experimental study, a eukaryotic expression vector containing
[
/pcDNA3.1(+)] was constructed and purified. ADMSCs were differentiated into IPCs. ADMSCs were assigned in two groups including control (C), and the
overexpressed (
+) groups. The ADMSCs were transfected by
/pcDNA 3.1(+) at day 10 of the differentiation. Differentiated cells were analyzed for the expression of multiple β cell specific genes (
,
,
,
,
,
, and
) using real-time polymerase chain reaction (PCR). The insulin secretion potency of the differentiated cells in response to glucose exposure was also determined using an enzyme-linked immunosorbent assay (ELISA) method and Dithizone (DTZ) staining. The IPCs from the control manipulated group, and un-differentiated ADMSCs group were transplanted to streptozotocin (STZ)-diabetic rats. Rats were monitored for blood glucose and insulin concentration.
The results revealed that ADMSCs were successfully differentiated into IPCs through the 14 day differentiation protocol. The expression of β-cell specific genes in
+ IPCs was higher than in control cells. Glucose-induced insulin secretion after the exposure of IPCs to glucose was higher in
+ group than the control group. The STZdiabetic rats showed an ability to secrete insulin and apparent hyperglycemic condition adjustment after transplantation of the control IPCs. The mean insulin concentration of diabetic rats that were transplanted by manipulated IPCs was significantly higher than ADMSCs-transplanted rats; however, no effect was observed in the concentration of bloodn glucose.
The overexpression of
can be used as a novel promising approach for the efficient production of IPCs from ADMSCs in vitro. However, the future therapeutic use of the
+ IPCs in diabetic animals needs further investigations.</description><identifier>ISSN: 2228-5806</identifier><identifier>EISSN: 2228-5814</identifier><identifier>DOI: 10.22074/cellj.2019.5669</identifier><identifier>PMID: 30825290</identifier><language>eng</language><publisher>Iran: Royan Institute of Iran</publisher><subject>Beta cells ; Blood ; Blood glucose ; Blood levels ; Cell differentiation ; Diabetes ; Diabetes mellitus ; Differentiation ; Enzyme-linked immunosorbent assay ; Gene expression ; Genes ; Glucose ; Insulin ; Insulin secretion ; Mesenchymal stem cells ; Nkx2.2 protein ; Nkx6.1 protein ; Original ; Polymerase chain reaction ; Stem cells ; Streptozocin ; Transplantation</subject><ispartof>Cell journal (Yakhteh), 2019-07, Vol.21 (2), p.169-178</ispartof><rights>Copyright© by Royan Institute. All rights reserved.</rights><rights>2019. This work is published under https://creativecommons.org/licenses/by-nc/3.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>The Cell Journal (Yakhteh) is an open access journal which means the articles are freely available online for any individual author to download and use the providing address.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6397604/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6397604/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27923,27924,53790,53792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30825290$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Dayer, Dian</creatorcontrib><creatorcontrib>Tabandeh, Mohammad Reza</creatorcontrib><creatorcontrib>Moghimipour, Eskandar</creatorcontrib><creatorcontrib>Hashemi Tabar, Mahmood</creatorcontrib><creatorcontrib>Ghadiri, AtaA</creatorcontrib><creatorcontrib>Allah Bakhshi, Elham</creatorcontrib><creatorcontrib>Orazizadeh, Mahmoud</creatorcontrib><creatorcontrib>Ghafari, Mohammad Ali</creatorcontrib><title>MafA Overexpression: A New Efficient Protocol for In Vitro Differentiation of Adipose-Derived Mesenchymal Stem Cells into Functional Insulin-Producing Cells</title><title>Cell journal (Yakhteh)</title><addtitle>Cell J</addtitle><description>We proposed a novel differentiation method for the efficient differentiation of adipose-derived mesenchymal stem cells (ADMSCs) into functional insulin-producing cells (IPCs) based on
overexpression.
In this experimental study, a eukaryotic expression vector containing
[
/pcDNA3.1(+)] was constructed and purified. ADMSCs were differentiated into IPCs. ADMSCs were assigned in two groups including control (C), and the
overexpressed (
+) groups. The ADMSCs were transfected by
/pcDNA 3.1(+) at day 10 of the differentiation. Differentiated cells were analyzed for the expression of multiple β cell specific genes (
,
,
,
,
,
, and
) using real-time polymerase chain reaction (PCR). The insulin secretion potency of the differentiated cells in response to glucose exposure was also determined using an enzyme-linked immunosorbent assay (ELISA) method and Dithizone (DTZ) staining. The IPCs from the control manipulated group, and un-differentiated ADMSCs group were transplanted to streptozotocin (STZ)-diabetic rats. Rats were monitored for blood glucose and insulin concentration.
The results revealed that ADMSCs were successfully differentiated into IPCs through the 14 day differentiation protocol. The expression of β-cell specific genes in
+ IPCs was higher than in control cells. Glucose-induced insulin secretion after the exposure of IPCs to glucose was higher in
+ group than the control group. The STZdiabetic rats showed an ability to secrete insulin and apparent hyperglycemic condition adjustment after transplantation of the control IPCs. The mean insulin concentration of diabetic rats that were transplanted by manipulated IPCs was significantly higher than ADMSCs-transplanted rats; however, no effect was observed in the concentration of bloodn glucose.
The overexpression of
can be used as a novel promising approach for the efficient production of IPCs from ADMSCs in vitro. However, the future therapeutic use of the
+ IPCs in diabetic animals needs further investigations.</description><subject>Beta cells</subject><subject>Blood</subject><subject>Blood glucose</subject><subject>Blood levels</subject><subject>Cell differentiation</subject><subject>Diabetes</subject><subject>Diabetes mellitus</subject><subject>Differentiation</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Glucose</subject><subject>Insulin</subject><subject>Insulin secretion</subject><subject>Mesenchymal stem cells</subject><subject>Nkx2.2 protein</subject><subject>Nkx6.1 protein</subject><subject>Original</subject><subject>Polymerase chain reaction</subject><subject>Stem cells</subject><subject>Streptozocin</subject><subject>Transplantation</subject><issn>2228-5806</issn><issn>2228-5814</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpdkUtv1DAUhS0EolXpnhWyxIZNBsfPmAXSaNrCSC1F4rGNHD9ajxI7tZ2B_hd-LK5aKsAbX-l-59xjXwBetmiFMRL0rbbjuFth1MoV41w-AYcY465hXUufPtaIH4DjnHeoHo5wK_FzcEBQhxmW6BD8ulBuDS_3Ntmfc7I5-xjewTX8ZH_AU-e89jYU-DnFEnUcoYsJbgP87kuK8MQ7V3WheFWqDEYH18bPMdvmxCa_twZe2GyDvr6d1Ai_FDvBTc2coQ8lwrMl6DtdbW1DXkYfmjrHLNqHq3vuBXjm1Jjt8cN9BL6dnX7dfGzOLz9sN-vzZsaSlkZpwgaEpDLEaer00A1GKclQJ2ohBMVkaCkTSDODuGgHqglyjFDT4bYzjByB9_e-8zJM1uj6pKTGfk5-Uum2j8r3_3aCv-6v4r7nRAqOaDV482CQ4s1ic-knn-_Wo4KNS-7rGMEwI7Kt6Ov_0F1cUv2ESnUUSc4kF5V69Xeixyh_Fkd-A-Dunu4</recordid><startdate>201907</startdate><enddate>201907</enddate><creator>Dayer, Dian</creator><creator>Tabandeh, Mohammad Reza</creator><creator>Moghimipour, Eskandar</creator><creator>Hashemi Tabar, Mahmood</creator><creator>Ghadiri, AtaA</creator><creator>Allah Bakhshi, Elham</creator><creator>Orazizadeh, Mahmoud</creator><creator>Ghafari, Mohammad Ali</creator><general>Royan Institute of Iran</general><general>Royan 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Mesenchymal Stem Cells into Functional Insulin-Producing Cells</title><author>Dayer, Dian ; Tabandeh, Mohammad Reza ; Moghimipour, Eskandar ; Hashemi Tabar, Mahmood ; Ghadiri, AtaA ; Allah Bakhshi, Elham ; Orazizadeh, Mahmoud ; Ghafari, Mohammad Ali</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p294t-ac35b009ad3fc4fcb8bdaa95087bda77423b14570c5d0671b4c30f534d8218d53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Beta cells</topic><topic>Blood</topic><topic>Blood glucose</topic><topic>Blood levels</topic><topic>Cell differentiation</topic><topic>Diabetes</topic><topic>Diabetes mellitus</topic><topic>Differentiation</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Gene expression</topic><topic>Genes</topic><topic>Glucose</topic><topic>Insulin</topic><topic>Insulin secretion</topic><topic>Mesenchymal stem cells</topic><topic>Nkx2.2 protein</topic><topic>Nkx6.1 protein</topic><topic>Original</topic><topic>Polymerase chain reaction</topic><topic>Stem cells</topic><topic>Streptozocin</topic><topic>Transplantation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dayer, Dian</creatorcontrib><creatorcontrib>Tabandeh, Mohammad Reza</creatorcontrib><creatorcontrib>Moghimipour, Eskandar</creatorcontrib><creatorcontrib>Hashemi Tabar, Mahmood</creatorcontrib><creatorcontrib>Ghadiri, AtaA</creatorcontrib><creatorcontrib>Allah Bakhshi, Elham</creatorcontrib><creatorcontrib>Orazizadeh, Mahmoud</creatorcontrib><creatorcontrib>Ghafari, Mohammad Ali</creatorcontrib><collection>PubMed</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Immunology Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology 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Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Cell journal (Yakhteh)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dayer, Dian</au><au>Tabandeh, Mohammad Reza</au><au>Moghimipour, Eskandar</au><au>Hashemi Tabar, Mahmood</au><au>Ghadiri, AtaA</au><au>Allah Bakhshi, Elham</au><au>Orazizadeh, Mahmoud</au><au>Ghafari, Mohammad Ali</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MafA Overexpression: A New Efficient Protocol for In Vitro Differentiation of Adipose-Derived Mesenchymal Stem Cells into Functional Insulin-Producing Cells</atitle><jtitle>Cell journal (Yakhteh)</jtitle><addtitle>Cell J</addtitle><date>2019-07</date><risdate>2019</risdate><volume>21</volume><issue>2</issue><spage>169</spage><epage>178</epage><pages>169-178</pages><issn>2228-5806</issn><eissn>2228-5814</eissn><abstract>We proposed a novel differentiation method for the efficient differentiation of adipose-derived mesenchymal stem cells (ADMSCs) into functional insulin-producing cells (IPCs) based on
overexpression.
In this experimental study, a eukaryotic expression vector containing
[
/pcDNA3.1(+)] was constructed and purified. ADMSCs were differentiated into IPCs. ADMSCs were assigned in two groups including control (C), and the
overexpressed (
+) groups. The ADMSCs were transfected by
/pcDNA 3.1(+) at day 10 of the differentiation. Differentiated cells were analyzed for the expression of multiple β cell specific genes (
,
,
,
,
,
, and
) using real-time polymerase chain reaction (PCR). The insulin secretion potency of the differentiated cells in response to glucose exposure was also determined using an enzyme-linked immunosorbent assay (ELISA) method and Dithizone (DTZ) staining. The IPCs from the control manipulated group, and un-differentiated ADMSCs group were transplanted to streptozotocin (STZ)-diabetic rats. Rats were monitored for blood glucose and insulin concentration.
The results revealed that ADMSCs were successfully differentiated into IPCs through the 14 day differentiation protocol. The expression of β-cell specific genes in
+ IPCs was higher than in control cells. Glucose-induced insulin secretion after the exposure of IPCs to glucose was higher in
+ group than the control group. The STZdiabetic rats showed an ability to secrete insulin and apparent hyperglycemic condition adjustment after transplantation of the control IPCs. The mean insulin concentration of diabetic rats that were transplanted by manipulated IPCs was significantly higher than ADMSCs-transplanted rats; however, no effect was observed in the concentration of bloodn glucose.
The overexpression of
can be used as a novel promising approach for the efficient production of IPCs from ADMSCs in vitro. However, the future therapeutic use of the
+ IPCs in diabetic animals needs further investigations.</abstract><cop>Iran</cop><pub>Royan Institute of Iran</pub><pmid>30825290</pmid><doi>10.22074/cellj.2019.5669</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6397604 |
| source | DOAJ Directory of Open Access Journals; PubMed Central Open Access; PubMed Central |
| subjects | Beta cells Blood Blood glucose Blood levels Cell differentiation Diabetes Diabetes mellitus Differentiation Enzyme-linked immunosorbent assay Gene expression Genes Glucose Insulin Insulin secretion Mesenchymal stem cells Nkx2.2 protein Nkx6.1 protein Original Polymerase chain reaction Stem cells Streptozocin Transplantation |
| title | MafA Overexpression: A New Efficient Protocol for In Vitro Differentiation of Adipose-Derived Mesenchymal Stem Cells into Functional Insulin-Producing Cells |
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