10x chromium single cell dissociation trypsin

In addition, we generated mouse single-cell transcriptomics data to contextualize our human findings with the mouse counterpart, facilitating translational research between these species. We uncover previously uncharacterized cellular heterogeneity in the somatic lineage, with relevance for gonadal conditions that have their origin during development, such as differences in sex development. Here, we used single-cell multiomics and spatial methods to disentangle the cellular and molecular programmes that mediate human gonadal development in space and time. Later in development, granulosa cells surround primary oocytes to form primordial follicles, remaining quiescent until puberty 4. In females, PGCs differentiate into oogonia, which enter an asynchronous transition from mitosis to meiosis. In males, primordial germ cells (PGCs), the gamete precursors, differentiate into prespermatogonia, forming cord-like structures with Sertoli cells and entering mitotic arrest.

Sertoli cells and preGCs coordinate the differentiation of the remaining sex-specific gonadal somatic (for example, interstitial) and germ cell lineages 3.

Around 6 weeks after conception (postconceptional weeks PCW) 1, gonadal somatic cells expressing SRY, the Y-linked testis-determining factor, differentiate into Sertoli cells (testicular supporting cells) or, in its absence, into pregranulosa cells (preGCs ovarian supporting cells) 2. In humans, the undifferentiated bipotent gonads, which emerge on the ventral surface of the mesonephros, commit to ovarian or testicular fate. This study provides a comprehensive spatiotemporal map of human and mouse gonadal differentiation, which can guide in vitro gonadogenesis. In males, we identify human SIGLEC15 + and TREM2 + fetal testicular macrophages, which signal to somatic cells outside and inside the developing testis cords, respectively.

In females, we resolve the cellular and molecular events that give rise to the first and second waves of granulosa cells that compartmentalize the developing ovary to modulate germ cell differentiation. In both species, we define the somatic cell states present at the time of sex specification, including the bipotent early supporting population that, in males, upregulates the testis-determining factor SRY and sPAX8s, a gonadal lineage located at the gonadal–mesonephric interface. We extracted human-specific regulatory programmes that control the development of germline and somatic cell lineages by profiling equivalent developmental stages in mice. Here, we generated a comprehensive map of first- and second-trimester human gonads using a combination of single-cell and spatial transcriptomics, chromatin accessibility assays and fluorescent microscopy. Historically, limited tissue accessibility, a lack of reliable in vitro models and critical differences between humans and mice have hampered our knowledge of human gonadogenesis, despite its importance in gonadal conditions and infertility. Gonadal development is a complex process that involves sex determination followed by divergent maturation into either testes or ovaries 1. Nature volume 607, pages 540–547 ( 2022) Cite this article Single-cell roadmap of human gonadal development

10x chromium single cell dissociation trypsin