Derivation and characterization of novel nonhuman primate embryonic stem cell lines from in vitro-fertilized baboon preimplantation embryos

Tiencheng A Chang, Ya-guang Liu, Carlton A. Eddy, Ethan S. Jacoby, Peter A. Binkley, Robert G. Brzyski, Robert S Schenken

Research output: Contribution to journalArticle

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Abstract

The development of nonhuman primate (NHP) embryonic stem cell (ESC) models holds great promise for cell-mediated treatment of debilitating diseases and to address numerous unanswered questions regarding the therapeutic efficacy of ESCs while supplanting ethical considerations involved with human studies. Here we report successful establishment and characterization of 3 novel baboon (Papio cynocephalus) ESC lines from the inner cell mass of intracytoplasmic sperm injection-derived blastocysts. Embryos were cultured in an improved baboon embryo in vitro culture protocol. The inner cell mass of blastocyst was laser-dissected and plated on mouse embryonic fibroblast feeder cell monolayer in the NHP ESC culture medium. Three cell lines with characteristic ESC morphology have been cultured through an extended period (>14 months), with 2 male cell lines (UT-1 and-2) and 1 female cell line (UT-3) displaying normal baboon karyotypes. Reverse transcription-polymerase chain reaction analysis confirmed that all 3 lines express primate ESC pluripotency markers, including OCT-4, NANOG, SOX-2, TERT, TDGF, LEFTYA, and REX-1. All 3 lines demonstrated positive immunocytochemical staining for OCT-4, stage-specific embryonic antigen-3, stage-specific embryonic antigen-4, TRA-1-60, and TRA-1-81. Baboon ESCs injected into NOD/SCID mice formed teratomas with all 3 germ layers. In addition, embryoid body-like spherical structures were derived and initial outgrowth was observed when embedded into extracellular matrix Matrigel ®. The ESC lines established in this NHP model have the potential to extend our knowledge in the fields of developmental biology, regenerative medicine, and future applications, including preclinical safety assessment of in vivo stem cell therapy.

Original languageEnglish (US)
Pages (from-to)1053-1062
Number of pages10
JournalStem Cells and Development
Volume20
Issue number6
DOIs
StatePublished - Jun 1 2011

Fingerprint

Papio
Blastocyst
Embryonic Stem Cells
Primates
Cell Line
Papio cynocephalus
Embryonic Structures
Blastocyst Inner Cell Mass
Embryoid Bodies
Feeder Cells
Germ Layers
Developmental Biology
Inbred NOD Mouse
Regenerative Medicine
SCID Mice
Intracytoplasmic Sperm Injections
Teratoma
Cell- and Tissue-Based Therapy
Karyotype
Reverse Transcription

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

Cite this

Derivation and characterization of novel nonhuman primate embryonic stem cell lines from in vitro-fertilized baboon preimplantation embryos. / Chang, Tiencheng A; Liu, Ya-guang; Eddy, Carlton A.; Jacoby, Ethan S.; Binkley, Peter A.; Brzyski, Robert G.; Schenken, Robert S.

In: Stem Cells and Development, Vol. 20, No. 6, 01.06.2011, p. 1053-1062.

Research output: Contribution to journalArticle

Chang, Tiencheng A ; Liu, Ya-guang ; Eddy, Carlton A. ; Jacoby, Ethan S. ; Binkley, Peter A. ; Brzyski, Robert G. ; Schenken, Robert S. / Derivation and characterization of novel nonhuman primate embryonic stem cell lines from in vitro-fertilized baboon preimplantation embryos. In: Stem Cells and Development. 2011 ; Vol. 20, No. 6. pp. 1053-1062.
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