Characterization of a novel embryonic stem cell line from an ICSI-derived blastocyst in the African green monkey

Abstract

Several cell types from the African green monkey (Cercopithecus aethiops), such as red blood cells, primary culture cells from kidney, and the Vero cell line, are valuable sources for biomedical research and testing. Embryonic stem (ES) cells that are established from blastocysts have pluripotency to differentiate into these and other types of cells. We examined an in vitro culture system of zygotes produced by ICSI in African green monkeys and attempted to establish ES cells. Culturing with and without a mouse embryonic fibroblast (MEF) cell monolayer resulted in the development of ICSI-derived zygotes to the blastocyst stage, while culturing with a buffalo rat liver cell monolayer yielded no development (3/14, 21.4% and 6/31, 19.4% vs 0/23, 0% respectively; P!0.05). One of the nine blastocysts, which had been one of the zygotes co-cultured with MEF cells, formed flat colonies consisting of cells with large nuclei, similar to other primate ES cell lines. The African green monkey ES (AgMES) cells expressed pluripotency markers, formed teratomas consisting of three embryonic germ layer tissues, and had a normal chromosome number. Furthermore, expression of the germ cell markers CD9 and DPPA3 (STELLA) was detected in the embryoid bodies, suggesting that AgMES cells might have the potential ability to differentiate into germ cells. The results suggested that MEF cells greatly affected the quality of the inner cell mass of the blastocysts. In addition, AgMES cells would be a precious resource for biomedical research such as other primate ES cell lines.

Derivation and Characterization of Novel Nonhuman Primate Embryonic Stem Cell Lines from In Vitro-Fertilized Baboon Preimplantation Embryos

“…A similar phenomenon of mixed expression patterns of pluripotency markers has been found in marmoset ESC lines. One study showed weak immunostaining with SSEA-3 and TRA-1-81 as com- African green monkey [12], cynomolgus monkey [12], as well as in a recently published study on olive baboon ESC lines [13], but negative in rhesus monkey ESC lines [10]. Another study showed that REX-1 is expressed in early passages of human ESC-derived fibroblast-like cells [36].…”

“…All demonstrated pluripotency by forming teratomas in SCID mice. SSEA-3 and À4 are not essential for human ESC pluripotency [32], and expression of these markers is vague in African green monkey ESC [12]. SSEA-3 expression is absent in cynomolgus monkey ESC [7], whereas both SSEA-3 and À4 are expressed in the rhesus monkey ESC [3,10].…”

“…In this study we found that the requirement of bFGF to maintain baboon ESC colonies in undifferentiated status is higher than it is indicated in human and other NHP ESC protocols. LIF has been widely used in primate ESC derivation and maintenance protocols [4,5,11,12], and later found less or not effective for maintaining undifferentiated status of ESC lines from human, cynomolgus, and rhesus monkey [7,10,34]. LIF receptor message is low or variable in human ESC lines compared to mouse ESC lines [34].…”

“…The first ESC lines were derived from mouse embryos [1], and this was followed by successful derivation of ESC lines in rhesus monkey (Macaca mulatta) [3], common marmoset (Callithrix jacchus) [4], and humans [5]. Later on, more ESC lines from rhesus monkey, common marmoset, and other nonhuman primate (NHP) species, including African green monkey (Cercopithecus aethiops), cynomolgus monkey (Macaca fascicularis), and, recently, olive baboon (Papio anubis), were also derived [6][7][8][9][10][11][12][13]. NHP ESCs hold great potential in biomedical research to address numerous unanswered questions regarding the regulatory mechanisms of primate ESCs, therapeutic efficacy, and application in regenerative medicine.…”

Transthyretin Amyloidosis in Aged Vervet Monkeys, as a Candidate for the Spontaneous Animal Model of Senile Systemic Amyloidosis

“…Although there is a US primate center with a vervet monkey colony (http://www.ncrr.nih.gov/comparative_medicine/resource_directory/primates.asp#verv), funds are insufficient for maintaining such a huge facility in Japan. Shimozawa et al (2010) developed embryonic stem cells from a vervet monkey, which may help reproduce them in the future. Now, captive vervet monkeys possess a novel raison d'etre under the field of biomedical science based on characteristics similar to human SSA.…”