Long‐Term Oocyte‐Like Cell Development in Cultures Derived from Neonatal Marmoset Monkey Ovary

Abstract

We use the common marmoset monkey (Callithrix jacchus) as a preclinical nonhuman primate model to study reproductive and stem cell biology. The neonatal marmoset monkey ovary contains numerous primitive premeiotic germ cells (oogonia) expressing pluripotent stem cell markers including OCT4A (POU5F1). This is a peculiarity compared to neonatal human and rodent ovaries. Here, we aimed at culturing marmoset oogonia from neonatal ovaries. We established a culture system being stable for more than 20 passages and 5 months. Importantly, comparative transcriptome analysis of the cultured cells with neonatal ovary, embryonic stem cells, and fibroblasts revealed a lack of germ cell and pluripotency genes indicating the complete loss of oogonia upon initiation of the culture. From passage 4 onwards, however, the cultured cells produced large spherical, free-floating cells resembling oocyte-like cells (OLCs). OLCs strongly expressed several germ cell genes and may derive from the ovarian surface epithelium. In summary, our novel primate ovarian cell culture initially lacked detectable germ cells but then produced OLCs over a long period of time. This culture system may allow a deeper analysis of early phases of female primate germ cell development and—after significant refinement—possibly also the production of monkey oocytes.

Small putative NANOG, SOX2, and SSEA-4-positive stem cells resembling very small embryonic-like stem cells in sections of ovarian tissue in patients with ovarian cancer

“…The observation of NANOG-positive primitive oocyte-like cells in “chambers” of cancerous ovaries, which very possibly developed from small putative stem cells, strongly expressing NANOG and are attached to them, confirms the previous observation on small putative stem cells in adult human ovaries that may develop into primitive oocyte-like cells in vitro, for example in the presence of follicular fluid rich on substances important for oocyte growth and maturation in humans [ 19 , 20 , 22 , 25 , 37 ] and in some other mammalian species such as mouse [ 26 ], pig [ 27 ], and marmoset monkey [ 77 ]. The development of stem cells into oocyte-like cells in vitro represents an important confirmation of the pluripotent character of stem cells, as experienced in embryonic stem cells (ESCs) [ 78 ] and human induced pluripotent stem cells (hiPSCs) [ 79 ].…”

The β-catenin/CBP-antagonist ICG-001 inhibits pediatric glioma tumorigenicity in a Wnt-independent manner

“…Library preparation and sequencing analysis were performed as described before [52, 53]. Candidate genes were filtered to a minimum of 2x fold changes and FDR-corrected p-value<0.05.…”

“…RNA was isolated from two independent biological replicates of KNS42 and UW479 cells, and three independent biological replicates of SF188 cells after treatment with ICG-001 or vehicle for 48h using the High Pure RNA Isolation Kit (Roche, Eisenach, Germany) according to manufacturer's instructions. Library preparation and sequencing analysis were performed as described before [ 52 , 53 ]. Candidate genes were filtered to a minimum of 2x fold changes and FDR-corrected p-value<0.05.…”

The β-catenin/CBP-antagonist ICG-001 inhibits pediatric glioma tumorigenicity in a Wnt-independent manner

“…RNA was isolated from two independent biological replicates of KNS42 and UW479 cells, and three independent biological replicates of SF188 cells after treatment with ICG-001 or vehicle for 48h using the High Pure RNA Isolation Kit (Roche, Eisenach, Germany) according to manufacturer's instructions. Library preparation and sequencing analysis were performed as described before [52,53]. Candidate genes were filtered to a minimum of 2x fold changes and FDR-corrected p-value<0.05.…”