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Publication

Interspecies Chimerism with Mammalian Pluripotent Stem Cells.

Cell | 2017

Interspecies blastocyst complementation enables organ-specific enrichment of xenogenic pluripotent stem cell (PSC) derivatives. Here, we establish a versatile blastocyst complementation platform based on CRISPR-Cas9-mediated zygote genome editing and show enrichment of rat PSC-derivatives in several tissues of gene-edited organogenesis-disabled mice. Besides gaining insights into species evolution, embryogenesis, and human disease, interspecies blastocyst complementation might allow human organ generation in animals whose organ size, anatomy, and physiology are closer to humans. To date, however, whether human PSCs (hPSCs) can contribute to chimera formation in non-rodent species remains unknown. We systematically evaluate the chimeric competency of several types of hPSCs using a more diversified clade of mammals, the ungulates. We find that naïve hPSCs robustly engraft in both pig and cattle pre-implantation blastocysts but show limited contribution to post-implantation pig embryos. Instead, an intermediate hPSC type exhibits higher degree of chimerism and is able to generate differentiated progenies in post-implantation pig embryos.

Pubmed ID: 28129541 RIS Download

Research resources used in this publication

Additional research tools detected in this publication

Salk Institute Razavi Newman Integrative Genomics and Bioinformatics Core Facility (IGC) (RRID:SCR_014842)

Antibodies used in this publication

Associated grants

Agency: NCI NIH HHS, United States
Id: P30 CA014195

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Salk Institute Razavi Newman Integrative Genomics and Bioinformatics Core Facility (IGC) (tool)

RRID:SCR_014842

Core facility established to assist the Salk community with integrating genomics data into their research. The primary focus of the core is to provide analysis support for next-generation sequencing applications.

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Oct3/4 Antibody (C-10) (antibody)

RRID:AB_628051

This monoclonal targets Oct-3/4

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Purified anti-Tubulin beta 3 (TUBB3) (antibody)

RRID:AB_10063408

This monoclonal targets Tubulin beta-3

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CD 31 Platelet Endothelial Cell Adhesion Molecule (PECAM) (antibody)

RRID:AB_394816

This monoclonal targets 129/Sv mouse-derived endothelioma cell line tEnd.1

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Rabbit Anti-Histone H4, trimethyl (Lys20) ChIP Grade Polyclonal Antibody, Unconjugated (antibody)

RRID:AB_306969

This polyclonal targets Histone H4 (tri methyl K20) - ChIP Grade

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H3K9me3-human,H3K9me3-mouse (antibody)

RRID:AB_306848

This polyclonal targets H3K9me3

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Mouse Anti-Actin, alpha-Smooth Muscle Monoclonal Antibody, Unconjugated, Clone 1A4 (antibody)

RRID:AB_262054

This monoclonal targets Actin, alpha, Smooth Muscle

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Ep-CAM (C-10) (antibody)

RRID:AB_627531

This monoclonal targets Human EPCAM

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Human Epithelial Antigen (antibody)

RRID:AB_2335685

This monoclonal targets MCF7 cell line

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Keratin, type II; cytokeratin 8/18; EndoA antibody - Brulet, P. / Kemler, R.; Institut Pasteur (antibody)

RRID:AB_531826

This monoclonal targets Keratin, type II; cytokeratin 8/18; EndoA

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GFP (antibody)

RRID:AB_591819

This polyclonal targets GFP

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pCXLE-EGFP (plasmid)

RRID:Addgene_27082

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FUW-tetO-hKLF4 (plasmid)

RRID:Addgene_20725

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FUW-tetO-hMYC (plasmid)

RRID:Addgene_20723

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pMD2.G (plasmid)

RRID:Addgene_12259

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pMDLg/pRRE (plasmid)

RRID:Addgene_12251

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FUW-M2rtTA (plasmid)

RRID:Addgene_20342

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pCXLE-hSK (plasmid)

RRID:Addgene_27078

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pCXLE-hUL (plasmid)

RRID:Addgene_27080

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pEGIP (plasmid)

RRID:Addgene_26777

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FUW-tetO-hOCT4 (plasmid)

RRID:Addgene_20726

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pCXLE-hOCT3/4-shp53-F (plasmid)

RRID:Addgene_27077

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FUW-tetO-hSOX2 (plasmid)

RRID:Addgene_20724

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pRSV-Rev (plasmid)

RRID:Addgene_12253

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CCD-1112Sk (cell line)

RRID:CVCL_2769

Cell line CCD-1112Sk is a Finite cell line with a species of origin Homo sapiens (Human)

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NOD.Cg-Prkdc^scid Il2rg/SzJ (organism)

RRID:IMSR_JAX:005557

Mus musculus with name NOD.Cg-Prkdc^scid Il2rg/SzJ from IMSR.

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C57BL/6J (organism)

RRID:IMSR_JAX:000664

Mus musculus with name C57BL/6J from IMSR.

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Benchling (web application)

RRID:SCR_013955

An online resource which helps researchers manage and organize labs and experimental results by supplying molecular biology software tools for experimental design and data analysis. Benchling provides tools for functions such as primer design and colony counting as well as CRISPR guide design and automated Gibson and Golden Gate cloning. Users can take notes in line with data, link data across entries, keep files and data in one place, and manage and keep track of team progress. An enterprise version of Benchling is available for scientists working within an organization with additional administrative, compliance, and security protocols.

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