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Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

ABSTRACT: Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

PROVIDER: PRJNA1057805 | ENA |

REPOSITORIES: ENA

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Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

Project description:The application of organoids has been limited by the lack of methods for producing uniformly mature organoids at scale. This study introduces an organoid culture platform, called UniMat, which addresses the challenges of uniformity and maturity simultaneously. UniMat is designed to not only ensure consistent organoid growth but also facilitate an unrestricted supply of soluble factors by a 3D geometrically-engineered, permeable membrane-based platform. Using UniMat, we demonstrate the scalable generation of kidney organoids with enhanced uniformity in both structure and function compared to conventional methods. Notably, kidney organoids within UniMat matured significantly better, showing increased expression of nephron transcripts, more in vivo-like cell-type balance, and better vascularization. Moreover, UniMat's design offers a more standardized organoid model for drug testing, as demonstrated by its consistent response to a polycystic-kidney-disease drug. In essence, UniMat presents a transformative platform for organoid technology, promising applications in organ development, disease modeling, and drug screening.

2024-09-07 | GSE252104 | GEO

Culture of mature adipocytes under a permeable membrane and comparative analysis with different cell culture models

Project description:White adipose tissue (WAT) is a central factor in the development of type 2 diabetes. Despite the epidemiological importance of WAT there is a paucity of translational models to study long term changes in mature adipocytes. Here, we describe a novel method for the culture of mature white adipocytes under a permeable membrane. Compared to existing culture methods such as adipose tissue explants and adipocyte ceiling culture, Membrane mature Adipocyte Aggregate Cultures (MAAC) are superior at maintaining adipogenic gene expression through 2 weeks of culture, do not dedifferentiate, and are under reduced hypoxic stress relative to adipose tissue explants. Unbiased RNA-Sequencing analysis indicates that the gene expression profile of MAAC in culture for 1 or 2 weeks is more similar to starting patient material than cultured adipose tissue explants, floating adipocytes, or in vitro-differentiated precursors from the same donor, with the fewest number of differentially expressed genes and the smallest fold-change in gene expression. Importantly, adipocytes from lean and obese patients can be cultured as MAAC, as can subcutaneous and visceral adipocytes, while maintaining depot-specific gene expression signatures. MAAC remain fully functional after long term culture, with similar responses to insulin and lipolytic stimuli compared to recently isolated cells. In addition, MAAC maintain the ability to crosstalk with other cell types, producing synergistic increases in IL6 and IL8 when co-cultured with macrophages, and respond robustly and predictably to diverse pharmacological agonism. Together, these abilities make MAAC a powerful tool for studying phenotypic changes in mature adipocytes, crosstalk between adipocytes and other cell types, and provide an improved translational model for drug development for the modulation of adipose tissue function.

2019-04-02 | GSE115020 | GEO

Scalable, methanol-free manufacturing of the SARS-CoV-2 receptor binding domain in engineered Komagataella phaffii

Project description:Prevention of COVID-19 on a global scale will require the continued development of high-volume, low-cost platforms for the manufacturing of vaccines to supply on-going demand. Vaccine candidates based on recombinant protein subunits remain important because they can be manufactured at low costs in existing large-scale production facilities that use microbial hosts like Komagataella phaffii (Pichia pastoris). Here, we report an improved and scalable manufacturing approach for the SARS-CoV-2 spike protein receptor binding domain (RBD); this protein is a key antigen for several reported vaccine candidates. We genetically engineered a manufacturing strain of K. phaffii to obviate the requirement for methanol-induction of the recombinant gene. Methanol-free production improved the secreted titer of the RBD protein by >5x by alleviating protein folding stress. Removal of methanol from the production process enabled scale up to a 1,200 L pre-existing production facility. This engineered strain is now used to produce an RBD-based vaccine antigen that is currently in clinical trials and could be used to produce other variants of RBD as needed for future vaccines.

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Uniform response to aneuploidy in human cells

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Scalable, methanol-free manufacturing of the SARS-CoV-2 receptor binding domain in engineered Komagataella phaffii

Project description:Scalable, methanol-free manufacturing of the SARS-CoV-2 receptor binding domain in engineered Komagataella phaffii

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Uniform response to aneuploidy in human cells

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