title: Interspecies Chimerism with Mammalian Pluripotent Stem Cells.
abstract: 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.
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Aida
authors.initials: A
authors.lastname: Platero-Luengo
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sakurai
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sugawara
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Maria Antonia
authors.initials: MA
authors.lastname: Gil
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Yamauchi
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Department of Animal Science, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Yanina Soledad
authors.initials: YS
authors.lastname: Bogliotti
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Cuello
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Mariana
authors.initials: M
authors.lastname: Morales Valencia
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Daiji
authors.initials: D
authors.lastname: Okumura
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Jingping
authors.initials: J
authors.lastname: Luo
authors.affiliation: Department of Animal Science, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Marcela
authors.initials: M
authors.lastname: Vilariño
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Parrilla
authors.affiliation: Department of Animal Science, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Delia Alba
authors.initials: DA
authors.lastname: Soto
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Cristina A
authors.initials: CA
authors.lastname: Martinez
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Hishida
authors.affiliation: Universidad Católica San Antonio de Murcia (UCAM) Campus de los Jerónimos, N° 135 Guadalupe 30107 Murcia, Spain.
authors.forename: Sonia
authors.initials: S
authors.lastname: Sánchez-Bautista
authors.affiliation: Universidad Católica San Antonio de Murcia (UCAM) Campus de los Jerónimos, N° 135 Guadalupe 30107 Murcia, Spain.
authors.forename: M Llanos
authors.initials: ML
authors.lastname: Martinez-Martinez
authors.affiliation: Department of Animal Science, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Huili
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Alicia
authors.initials: A
authors.lastname: Nohalez
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Emi
authors.initials: E
authors.lastname: Aizawa
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Paloma
authors.initials: P
authors.lastname: Martinez-Redondo
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Ocampo
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Reddy
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Roca
authors.affiliation: Department of Animal Science, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Maga
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Concepcion Rodriguez
authors.initials: CR
authors.lastname: Esteban
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: W Travis
authors.initials: WT
authors.lastname: Berggren
authors.affiliation: Universidad Católica San Antonio de Murcia (UCAM) Campus de los Jerónimos, N° 135 Guadalupe 30107 Murcia, Spain.
authors.forename: Estrella
authors.initials: E
authors.lastname: Nuñez Delicado
authors.affiliation: Universidad Católica San Antonio de Murcia (UCAM) Campus de los Jerónimos, N° 135 Guadalupe 30107 Murcia, Spain.
authors.forename: Jeronimo
authors.initials: J
authors.lastname: Lajara
authors.affiliation: Clinica Centro Fundación Pedro Guillén, Clínica CEMTRO, Avenida Ventisquero de la Condesa 42, 28035 Madrid, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Guillen
authors.affiliation: Universidad Católica San Antonio de Murcia (UCAM) Campus de los Jerónimos, N° 135 Guadalupe 30107 Murcia, Spain; Clinica Centro Fundación Pedro Guillén, Clínica CEMTRO, Avenida Ventisquero de la Condesa 42, 28035 Madrid, Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Guillen
authors.affiliation: Hospital Clínico de Barcelona-IDIBAPS, Universitat de Barcelona, 08007 Barcelona, Spain.
authors.forename: Josep M
authors.initials: JM
authors.lastname: Campistol
authors.affiliation: Department of Animal Medicine and Surgery, University of Murcia Campus de Espinardo, 30100 Murcia, Spain.
authors.forename: Emilio A
authors.initials: EA
authors.lastname: Martinez
authors.affiliation: Department of Animal Science, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Pablo Juan
authors.initials: PJ
authors.lastname: Ross
authors.affiliation: Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA. Electronic address: belmonte@salk.edu.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Izpisua Belmonte
mesh_terms: D000818:Animals; D001755:Blastocyst; D064113:CRISPR-Cas Systems; D002417:Cattle; D046528:Chimerism; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D000072669:Gene Editing; D006801:Humans; D008297:Male; D008322:Mammals / Q000145:classification / Q000196:embryology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D039904:Pluripotent Stem Cells; D051381:Rats; D017207:Rats, Sprague-Dawley; D034421:Sus scrofa
keywords: CRISPR-Cas9; human naïve pluripotent stem cells; human-cattle chimeric embryo; human-pig chimeric embryo; interspecies blastocyst complementation; interspecies chimera; organ and tissue generation; pluripotent stem cells; zygote genome editing