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1.
Langmuir ; 2024 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-39137165

RESUMO

Traditional screen printing is an easy approach commonly used for conductive pattern fabrication of electronics but lacks high resolution. Photolithography offers better resolution but is complex. Photosensitive silver pastes (PSP) combine the benefits of both but suffer from undercut issues, causing uneven etching, decreased interfacial adhesion, and thus poor resolutions. In this study, we explore the use of molecular precursors (i.e., silver oxalate) to replace metallic silver particles and enhance the depth of light penetration. Our findings demonstrate a successful solution to the undercut issue, achieving an undercut index of 1.0, indicating an undercut-free scenario and enabling higher resolutions in line and pattern formation. Additionally, our research confirms the feasibility of multilayer stacking of photosensitive pastes, achieving unprecedented aspect ratios in line patterns. By replacing 25% of micrometer silver powder with silver oxalate (PSP-25), we achieved optimal line widths as fine as 10 µm. The three-layer stack of PSP-25 reached a substantial aspect ratio with a height of 29.4 µm and an optimal fringe pattern resolution of 10 µm line width with a 15 µm aisle width. Utilization of silver oxalate was observed to slightly expand the line width, likely due to light scattering by the fine silver nanoparticles (∼40 nm) formed during the photodecomposition of silver oxalate.

2.
Cell Stem Cell ; 31(6): 921-939.e17, 2024 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-38692273

RESUMO

Nephron progenitor cells (NPCs) self-renew and differentiate into nephrons, the functional units of the kidney. Here, manipulation of p38 and YAP activity allowed for long-term clonal expansion of primary mouse and human NPCs and induced NPCs (iNPCs) from human pluripotent stem cells (hPSCs). Molecular analyses demonstrated that cultured iNPCs closely resemble primary human NPCs. iNPCs generated nephron organoids with minimal off-target cell types and enhanced maturation of podocytes relative to published human kidney organoid protocols. Surprisingly, the NPC culture medium uncovered plasticity in human podocyte programs, enabling podocyte reprogramming to an NPC-like state. Scalability and ease of genome editing facilitated genome-wide CRISPR screening in NPC culture, uncovering genes associated with kidney development and disease. Further, NPC-directed modeling of autosomal-dominant polycystic kidney disease (ADPKD) identified a small-molecule inhibitor of cystogenesis. These findings highlight a broad application for the reported iNPC platform in the study of kidney development, disease, plasticity, and regeneration.


Assuntos
Néfrons , Organoides , Animais , Organoides/citologia , Organoides/metabolismo , Humanos , Néfrons/citologia , Camundongos , Diferenciação Celular , Células-Tronco Pluripotentes Induzidas/metabolismo , Células-Tronco Pluripotentes Induzidas/citologia , Podócitos/metabolismo , Podócitos/citologia , Rim/patologia , Rim Policístico Autossômico Dominante/patologia , Rim Policístico Autossômico Dominante/metabolismo , Rim Policístico Autossômico Dominante/genética , Modelos Biológicos , Edição de Genes
3.
bioRxiv ; 2023 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-37293038

RESUMO

Nephron progenitor cells (NPCs) self-renew and differentiate into nephrons, the functional units of the kidney. Here we report manipulation of p38 and YAP activity creates a synthetic niche that allows the long-term clonal expansion of primary mouse and human NPCs, and induced NPCs (iNPCs) from human pluripotent stem cells. Cultured iNPCs resemble closely primary human NPCs, generating nephron organoids with abundant distal convoluted tubule cells, which are not observed in published kidney organoids. The synthetic niche reprograms differentiated nephron cells into NPC state, recapitulating the plasticity of developing nephron in vivo. Scalability and ease of genome-editing in the cultured NPCs allow for genome-wide CRISPR screening, identifying novel genes associated with kidney development and disease. A rapid, efficient, and scalable organoid model for polycystic kidney disease was derived directly from genome-edited NPCs, and validated in drug screen. These technological platforms have broad applications to kidney development, disease, plasticity, and regeneration.

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