# Dissect formative pluripotency using cultured pluripotent stem cells

> **NIH NIH R01** · UT SOUTHWESTERN MEDICAL CENTER · 2022 · $340,568

## Abstract

PROJECT SUMMARY/ABSTRACT
 Derivation of pluripotent stem cells (PSCs) has revolutionized developmental biology and regenerative
medicine. To stably maintain PSCs in culture and guide them to differentiate with high efficiency and fidelity into
a variety of cell types, it is important to understand the molecular mechanisms governing pluripotency (the ability
of a cell to generate any tissues in the body). Two phases of pluripotency, naïve and primed, have been defined
and studied in detail thanks to the successful derivation of mouse embryonic stem cells (ESCs) and epiblast
stem cells (EpiSCs), respectively. Mouse ESCs most closely resemble epiblast from a 4-day-old mouse
blastocyst (~embryonic day 4, or E4), while “primed” EpiSCs display a global gene expression signature similar
to the E7 epiblast of a post-implantation mouse embryo. Despite these advances, however, however, there is
lack of a well-established PSC model that resembles E5-6 early post-implantation epiblast, which corresponds
to the formative phase of pluripotency. Formative pluripotency exists within a time window during which naïve
pluripotency is reconfigured to prepare for multilineage competency, including germ cells. Functionally, formative
pluripotency is characterized by both chimera competency and permissiveness for direct primordial germ cell
(PGC) induction. Several recent studies have attempted to define this state by transient epiblast-like cells
(EpiLCs) differentiated from ESCs. To date, however, stable formative PSCs have not yet been generated. By
modulating the FGF, TGF-β and WNT pathways, we recently derived PSCs from both mice and humans (referred
to as FTW-PSCs) that are permissive for direct PGC-like cell induction in vitro and are capable of contributing to
intra- or inter-species chimeras in vivo. FTW-PSCs harbor molecular, cellular and phenotypic features
characteristic of formative pluripotency. The overall objective of this proposal is to use these newly
established cell lines to comprehensively dissect the formative state across species. The proposed
studies will elucidate the roles of several transcription factors in regulating mouse and human formative
pluripotency, as well as demonstrate that FTW-PSCs are a robust platform for dissecting the molecular
mechanisms underlying human and mouse PGC specification. In addition, we will establish an in vitro platform
for the generation of functional mouse oocytes and human oogonia based on formative FTW-PSCs, thereby
providing an invaluable resource for studying germ cell development and human infertility. Our proposal has
tremendous potential to revolutionize regenerative medicine and reproductive biology.

## Key facts

- **NIH application ID:** 10455585
- **Project number:** 5R01GM138565-02
- **Recipient organization:** UT SOUTHWESTERN MEDICAL CENTER
- **Principal Investigator:** Jun Wu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $340,568
- **Award type:** 5
- **Project period:** 2021-07-22 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10455585

## Citation

> US National Institutes of Health, RePORTER application 10455585, Dissect formative pluripotency using cultured pluripotent stem cells (5R01GM138565-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10455585. Licensed CC0.

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