Volume 16, Issue 3, May 2016, Pages 617–621 Generation of iPSC from cardiac and tail-tip fibroblasts derived from a second heart field reporter mousea b c Received 26 February 2016, Revised 9 March 2016, Accepted 17 March 2016, Available online 19 March 2016Mef2c Anterior Heart Field (AHF) enhancer is activated during embryonic heart development and it is expressed in multipotent cardiovascular progenitors (CVP) giving rise to endothelial and myocardial components of the outflow tract, right ventricle and ventricular septum. Here we have generated iPSC from transgenic Mef2c-AHF-Cre x Ai6(RCLZsGreen) mice. These iPSC will provide a novel tool to investigate the AHF-CVP and their cell progeny.Resource TableName of Stem Cell lineAHFiPS7 and AHFiPS19InstitutionCell Therapy Program, Foundation for Applied Medical Research (FIMA), University of Navarra.Person who created resourceXonia Carvajal-Vergara, Estibaliz Arellano-Viera, Javier LinaresContact person and emailxcarvajal@unav.es; fprosper@unav.esDate archived/stock dateMarch 2015OriginMouse tail-tip fibroblasts and mouse cardiac fibroblastsType of resourceBiological reagent: mouse induced pluripotent stem cell (iPSC) lines derived from transgenic Mef2c-AHF-Cre x Ai6(RCLZsGreen) miceSub-typeMouse induced pluripotent stem cell (iPSC) linesKey transcription factorsOct4, Sox2, cMyc, Klf4AuthenticationIdentity and purity of cell line confirmed (Fig. 1 ;
Fig. 2)Link to related literatureNot availableInformation in public databasesNot availableFull-size tableResource DetailsTail-tip fibroblasts (TTF) and cardiac fibroblasts (CF) derived from Mef2c-AHF-Cre (Verzi et al., 2005) x Ai6(RCLZsGreen) (Madisen et al., 2010) mice were transduced with Oct4, Sox2, Klf4 and c-Myc (OSKM) with the pMXs retrovirus vectors. Mouse ES-like colonies appeared 15–20 days after transduction and were picked and seeded on irradiated mouse embryonic fibroblasts (ɣMEFs) and maintained with LIF. Established iPSC, AHF-iPSC, derived from TTF (AHFiPS7) and CF (AHFiPS19) encoded the expected genomic insertions (Fig. 1a). The karyotypes of AHF-iPSC were normal (Fig. 1b) and the transgenes were silenced in established AHF-iPSC lines as shown by qRT-PCR (Fig. 1c). Endogenous pluripotency-associated markers such Oct4, Sox2, Nanog and Zfp42 were expressed in AHF-iPSC, analyzed by qRT-PCR (Fig. 1d), and Nanog (Fig. 1e) and alkaline phosphatase (AP, Fig. 1f) expression was verified by cell staining.
To demonstrate the capacity of AHF-iPSC lines to differentiate into the three germ layers we performed in vivo teratoma and in vitro embryoid body (EB) differentiation assays. Teratomas contained tissues derived from the three germ layers (Fig. 2a). We collected RNA from undifferentiated AHF-iPSC and from EB on day 7 and 14 of differentiation. Increased expression of Cxcl12/Mash1 (ectoderm), Acta2/Myh6 (mesoderm) and Hnf4a/Afp (endoderm), (Fig. 2b) was observed. Mef2c and ZsGreen were not detected in undifferentiated AHF-iPSC, however, the expression of Mef2c and ZsGreen was confirmed by qRT-PCR on day 7 and 14 of differentiation (Fig. 2b), and ZsGreen was observed under fluorescence microscopy (Fig. 2c).reseed Also found in: Thesaurus. reseed () vb (tr) Legend:Switch to new thesaurus Verb1.reseed - seed again or anewfarming, husbandry, agriculture - the practice of cultivating the land or raising stockseed - go to seed; "The dandelions went to seed"2.reseed - maintain by seeding without human intervention;
"Some plants reseed themselves indefinitely"maintain, sustain, keep - supply with necessities and support; "She alone sustained her family"; "The money will sustain our good cause"; "There's little to earn and many to keep" reseed vt [+ lawn, pitch] → refaire References in periodicals archive ▲research quantumresearch rocketresearch staffresearch studentresearch workresearch workerresearchedresearcherresearchfulresearchistreseasonreseatreseauresectresectabilityresectableresectionresectoscoperesectoscopyresecureresedaReseda luteolaReseda odorataResedaceaereseereseedreseekresegregateresegregationreseizeReseizerreseizurereselectreselectionresellresellerResemblableresemblanceresemblantresembleresemblerResemblinglyReseminateresendresensitizeresentresentencere-sentencingresenterresentfulresentfully▼ ReSecurity First Legal AssociationOnline Checking of Driving Licence Status Checking driving licence validity is easy with the online service offered by the Transport Department.
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