Dataset Information

Identify the protein contents within the fluids secreted by transplanted human vascularized lung organoid

ABSTRACT: We recently established a hPSC-derived 3D organoid system to mimic the vascularization process during lung differentiation. To further mature the organoid, we embedded them into the mouse kidney capsule for three months and observed the accumulation of fluid within the growing organoids. Therefore, we aim to understand if there are any proteins related to lung function are being secreted in this case.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Minzhe Guo

LAB HEAD: Mingxia Gu

PROVIDER: PXD048316 | Pride | 2025-07-06

REPOSITORIES: Pride

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Co-development of mesoderm and endoderm enables organotypic vascularization in lung and gut organoids.

Miao Yifei Y Pek Nicole M NM Tan Cheng C Jiang Cheng C Yu Zhiyun Z Iwasawa Kentaro K Shi Min M Kechele Daniel O DO Sundaram Nambirajan N Pastrana-Gomez Victor V Sinner Debora I DI Liu Xingchen X Lin Ko Chih KC Na Cheng-Lun CL Kishimoto Keishi K Yang Min-Chi MC Maharjan Sushila S Tchieu Jason J Whitsett Jeffrey A JA Zhang Yu Shrike YS McCracken Kyle W KW Rottier Robbert J RJ Kotton Darrell N DN Helmrath Michael A MA Wells James M JM Takebe Takanori T Zorn Aaron M AM Chen Ya-Wen YW Guo Minzhe M Gu Mingxia M

Cell 20250627

The vasculature and mesenchyme exhibit distinct organ-specific characteristics adapted to local physiological needs, shaped by microenvironmental and cell-cell interactions from early development. To recapitulate this entire process, we co-differentiated mesoderm and endoderm within the same spheroid to vascularize lung and intestinal organoids from induced pluripotent stem cells (iPSCs). Bone morphogenetic protein (BMP) signaling fine-tuned the endoderm-to-mesoderm ratio, a critical step in gen ...[more]

PMID: 40592324

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Project description:Underdeveloped lungs are the primary cause of death in premature infants, however, little is known about stem and progenitor cell maintenance during human lung development. In this study, we have identified that FGF7, Retinoic Acid and CHIR-99021, a small molecule that inhibits GSK3 to activate Wnt signaling, support in vitro maintenance of primary human fetal lung bud tip progenitor cells in a progenitor state. Furthermore, these factors are sufficient to derive a population of human bud tip-like progenitor cells in 3D organoid structures from human pluripotent stem cells (hPSC). Functional studies showed that hPSC-derived bud tip progenitor organoids do not contain any mesenchymal cell types, maintain multilineage potential in vitro and are able to engraft into the airways of injured mice and respond to systemic factors. We performed RNA-sequencing to assess the degree of similarity in global gene expression profiles between the full human fetal lung (59-127 days gestation), isolated human fetal bud tip progenitors, organoids grown from primary fetal bud tip progenitors, and hPSC-derived bud tip organoids. Results showed that hPSC-derived organoids have molecular profiles similar to organoids generated from primary human fetal lung tissue. Gene expression differences between hPSC-derived bud tip organoids and fetal progenitor organoids may be related to the presence of contaminating mesenchymal cells in primary cultures. hPSC-derived bud tip organoids are generated from a well-defined human cell sources, offering a distinct advantage over rare primary tissue as a means to study human specific lung development, homeostasis and disease. Sample Nomenclature - Description ------------------------------------------------------------------------- Peripheral fetal lung the distal/peripheral portion of the fetal lung (i.e., distal 0.5 cm) was excised from the rest of the lung using a scalpel. This includes all components of the lung (e.g., epithelial, mesenchymal, vascular). Isolated fetal bud tip the bud peripheral portion of the fetal lung was excised with a scalpel and subjected to enzymatic digestion and microdissection. The epithelium was dissected and separated from the mesenchyme, but a small amount of associated mesenchyme likely remained. Fetal progenitor organoid 3D organoid structures that arose from culturing isolated fetal epithelial bud tips. Foregut spheroid 3D foregut endoderm structure as described in Dye et al. (2015). Gives rise to patterned lung organoid (PLO) when grown in 3F medium. Patterned lung organoid (PLO) lung organoids that were generated by differentiating hPSCs, as described throughout the manuscript. Bud tip organoid organoids derived from PLOs, enriched for SOX2/SOX9 co-expressing cells, and grown/passaged in 3F medium.

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