Establishment and characterization of baboon embryonic stem cell lines: An Old World Primate model for regeneration and transplantation research

Abstract

Here we have developed protocols using the baboon as a complementary alternative Old World Primate to rhesus and other macaques which have severe limitations in their availability. Baboons are not limited as research resources, they are evolutionarily closer to humans and the multiple generations of pedigreed colonies which display complex human disease phenotypes all support their further optimization an invaluable primate model. Since neither baboon assisted reproductive technologies nor baboon embryonic stem cells (ESCs) have been reported, here we describe the first derivations and characterization of baboon ESC lines from IVF-generated blastocysts. Two ESCs lines (BabESC-4 and BabESC-15) display ESC morphology, express pluripotency markers (Oct-4, hTert, Nanog, Sox-2, Rex-1, TRA1–60, TRA1–81), and maintain stable euploid female karyotypes with parentage confirmed independently. They have been grown continuously for >430 and 290 days, respectively. Teratomas from both lines have all three germ layers. Availabilities of these BabESCs represent another important resource for stem cell biologists.

Derivation of Induced Pluripotent Stem Cells from the Baboon: A Nonhuman Primate Model for Preclinical Testing of Stem Cell Therapies

“…Colony morphology appeared similar among lines (Fig. 1A-D) and resembled that reported for human iPSCs (Takahashi et al, 2007;Yu et al, 2007), human ESCs (Thomson et al, 1998), and baboon ESCs (Simerly et al, 2009), including large flat colonies, with welldefined borders and a high nuclear/cytoplasm ratio.…”

“…Pluripotent (ESCs and/or iPSCs) stem cells have now been derived from at least six different NHP species: Rhesus monkeys (ESCs and iPSCs; Chan et al, 2011;Liu et al, 2008;Mitalipov et al, 2006;Navara et al, 2007;Pau and Wolf 2004;Thomson et al, 1995), marmosets (ESCs and iPSCs; Muller et al, 2009;Sasaki et al, 2005;Thomson et al, 1996;Wu et al, 2010), cynomolgus monkeys (ESCs and iPSCs; Suemori and Nakatsuji 2006;Suemori et al, 2001), pigtail macaques (iPSCs; Gori et al, 2012), drills (iPSCs; Ben-Nun et al, 2011), and baboons (ESCs, Chang et al, 2011;Simerly et al, 2009). Here we describe the first derivation of iPSCs from the baboon.…”

“…Derivation of baboon ESCs was previously accomplished by Simerly et al (2009). These authors noted several advantages of the baboon as a model NHP system for studies of stem cell-based therapeutic applications.…”

Ex Vivo Reconstitution of Arterial Endothelium by Embryonic Stem Cell-Derived Endothelial Progenitor Cells in Baboons

“…The BAB15 baboon ESC line at passage 39 was used in this study. They were cultured according to the published method and used within no more than 10 passages [28]. Undifferentiated ESCs were maintained with feeder cells in the ESC medium containing 15% knockout serum replacement; 1% nonessential amino acids (100 · ); 1% l-glutamine (100 · ); 1% antibiotic-antimycotic (100 · ); 100 mM b-mercaptoethanol; and 12 ng/mL human basic fibroblast growth factor (FGF) in knockout Dulbecco modified Eagle's minimal essential medium (DMEM).…”

Techniques and Conditions for Embryonic Germ Cell Derivation and Culture

“…Thus, it seems reasonable to suggest that questions regarding the process of reprogramming and differentiation via genomic imprints could potentially be more easily and appropriately addressed using non-human primate EGCs. To date, there are no reports on the isolation of stable long term EGC lines from non-human primate PGCs, but short term EGCs have been developed from the groups of Schatten and Simerly (Simerly et al, 2010). Baboon embryos were collected between days 28-35 post coitum (similar to human Carnegie Stages 12-16), dissecting the lower half of the embryo and excising the genital ridge, dorsal mesentery and part of the gut.…”