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Uncovering human yolk sac-derived innate lymphoid-biased multipotent progenitors prior to hematopoietic stem cell formation (CITE-Seq)

ABSTRACT: Uncovering human yolk sac-derived innate lymphoid-biased multipotent progenitors prior to hematopoietic stem cell formation (CITE-Seq)

PROVIDER: PRJNA1114754 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29131457_1.fastq.gz	Fastqsanger.gz
	SRR29131457_2.fastq.gz	Fastqsanger.gz
	SRR29131458.fastq.gz	Fastqsanger.gz

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Uncovering human yolk sac-derived innate lymphoid-biased multipotent progenitors prior to hematopoietic stem cell formation (CITE-Seq)

Project description:Hematopoietic stem cell (HSC)-independent lymphopoiesis has been elucidated in murine embryos. However, our understanding regarding human embryonic counterparts remains limited. Here, we unveiled the presence of human yolk sac-derived lymphoid-biased progenitors (YSLPs) expressing CD34, IL7R, LTB and IRF8 at Carnegie Stage 10, much earlier than the first HSCs emergence. The number and lymphopoietic potential of these progenitors were both significantly higher in yolk sac than embryo proper at this early stage. Importantly, single-cell/bulk culture and CITE-seq have elucidated the tendency of YSLP differentiating into innate lymphoid cells and dendritic cells. Notably, lymphoid progenitors in fetal liver before and after HSCs seeding displayed distinct transcriptional features, with the former closely resembling those of YSLPs.

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Expression data from Tnrc6a (GW182) mutant yolk sac

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Expression data from Tnrc6a (GW182) mutant yolk sac

Project description:GW182 (Tnrc6a) is a key component of RISC (miRNA-Induced Silencing Complex) that plays a critical role in miRNA-mediated gene silencing. Here, we show that GW182 is expressed in the yolk sac endoderm, and that gene-trap disruption of GW182 leads to growth arrest of yolk sac endoderm, impaired hematopoiesis and embryonic lethality. To investigate roles of GW182 in the yolk sac endoderm, we assessed changes in mRNA expression in the yolk sac of E9.5 GW182gt/gt embryos using microarrays (Affymetrix). Yolk sac of wild type littermates and GW182gt/gt embryos at E9.5 was collected for total RNA isolation using Trizol (Invitrogen). RNAs were purified according to the manufacturer’s protocol before subjected to Mouse Gene 1.0 ST Whole Genome Array (Affymetrix) for mRNA expression profiling. Experiments were performed in triplicate. Differentially expressed mRNAs were identified using a two-sample t-test (P<0.05 considered significant).

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Project description:The human definitive yolk sac is an important organ supporting the early developing embryo through nutrient supply and by facilitating the establishment of the embryonic circulatory system. However, the molecular and cellular biology of the human yolk sac remains largely obscure due to the lack of suitable in vitro models. Here, we show that human induced pluripotent stem cells (hiPSCs) co-cultured with various types of stromal cells as spheroids self-organize into yolk sac-like organoids without the addition of exogenous factors. Yolk sac-like organoids recapitulated a yolk sac specific cellular complement and structures as well as the functional ability to generate definitive hematopoietic progenitor cells (HPCs). Furthermore, sequential hemato-vascular ontogenesis could be observed during organoid formation. Notably, our organoid system can be performed in a scalable, autologous, and xeno-free condition, thereby providing an important model of human definitive yolk sac development and allows for efficient bulk generation of hiPSC-derived HPCs.

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