Biology subjects

Wolf, K. J.

Publications and source records attributed to Wolf, K. J..

4 recordsLinked to original sources

bioRxiv2026

Scalable generation of human stem cell-derived ureteric bud and collecting duct organoids in stirred bioreactors

Emerging efforts to engineer multicellular kidney tissues for drug discovery, disease modeling, and therapeutic repair require scalable generation of kidney organoids derived from human induced pluripotent stem cells (hiPSCs). Here, we report a method for generating kidney ureteric bud and collecting duct epithelial organoids in stirred tank bioreactors. We demonstrate that extracellular matrix microcarriers facilitate organoid expansion and are compatible with suspension bioreactor methods. The combination of cell-coated matrix microcarriers with stirred bioreactors promotes maturation marker expression compared to static controls comprising matrix-embedded organoids. Our method offers promise for generating epithelial organoids that serve as multicellular models and organ building blocks for scalable biofabrication.

bioengineering↗

bioRxiv2025

Perfusable 3D models of ureteric bud and collecting duct tubules

Recent protocols have emerged to derive ureteric bud (UB) and collecting duct (CD) organoids directly from human induced pluripotent stem cells (hiPSCs). However, these 3D kidney tissues lack biophysical cues from luminal flow and a drainage outlet. To address these limitations, we have created perfusable 3D models of UB and CD tubules. UB organoids are first generated from hiPSCs followed by their dissociation into individual UB cells. Individual UB cells are then seeded onto a 3D perfusable channel embedded within an extracellular matrix composed of fragmented basement membrane matrix and collagen I, where they self-assemble into a confluent monolayer. During in vitro perfusion, these cells exhibit UB-like marker expression over several weeks, during which they undergo budding akin to early branching morphogenesis in developing kidneys. To further promote network formation, UB cells are bioprinted adjacent to a perfusable UB tubule, which form interconnections through luminal fusion. Finally, these 3D perfusable UB tubules are differentiated into collecting duct tubules under luminal flow. Our platform facilitates fundamental understanding of human collecting duct formation during renal development, while paving the way for using these physiologically relevant models for drug testing, disease modeling, and, ultimately, integration into bioprinted kidney tissues for therapeutic use.

bioengineering↗

bioRxiv2023

Multi-omic screening of invasive GBM cells in engineered biomaterials and patient biopsies reveals targetable transsulfuration pathway alterations

While the poor prognosis of glioblastoma arises from the invasion of a subset of tumor cells, little is known of the metabolic alterations within these cells that fuel invasion. We integrated spatially addressable hydrogel biomaterial platforms, patient site-directed biopsies, and multi-omics analyses to define metabolic drivers of invasive glioblastoma cells. Metabolomics and lipidomics revealed elevations in the redox buffers cystathionine, hexosylceramides, and glucosyl ceramides in the invasive front of both hydrogel-cultured tumors and patient site-directed biopsies, with immunofluorescence indicating elevated reactive oxygen species (ROS) markers in invasive cells. Transcriptomics confirmed upregulation of ROS-producing and response genes at the invasive front in both hydrogel models and patient tumors. Amongst oncologic ROS, hydrogen peroxide specifically promoted glioblastoma invasion in 3D hydrogel spheroid cultures. A CRISPR metabolic gene screen revealed cystathionine gamma lyase (CTH), which converts cystathionine to the non-essential amino acid cysteine in the transsulfuration pathway, to be essential for glioblastoma invasion. Correspondingly, supplementing CTH knockdown cells with exogenous cysteine rescued invasion. Pharmacologic CTH inhibition suppressed glioblastoma invasion, while CTH knockdown slowed glioblastoma invasion in vivo. Our studies highlight the importance of ROS metabolism in invasive glioblastoma cells and support further exploration of the transsulfuration pathway as a mechanistic and therapeutic target.

cancer biology↗

bioRxiv2018

A 3D Topographical Model of Parenchymal Infiltration and Perivascular Invasion in Glioblastoma

Glioblastoma (GBM) is the most common and invasive primary brain cancer. GBM tumors are characterized by diffuse infiltration, with tumor cells invading slowly through the hyaluronic acid (HA)-rich parenchyma toward vascular beds and then migrating rapidly along microvasculature. Progress in understanding local infiltration, vascular homing, and perivascular invasion is limited by an absence of culture models that recapitulate these hallmark processes. Here we introduce a platform for GBM invasion consisting of a tumor-like cell reservoir and a parallel open channel \"vessel\" embedded in 3D HA-RGD matrix. We show that this simple paradigm is sufficient to capture multi-step invasion and transitions in cell morphology and speed reminiscent of those seen in GBM. Specifically, seeded tumor cells grow into multicellular masses that expand and invade the surrounding HA-RGD matrices while extending long (10-100 {micro}m), thin protrusions resembling those observed for GBM in vivo. Upon encountering the channel, cells orient along the channel wall, adopt a 2D-like morphology, and migrate rapidly along the channel. Structured illumination microscopy reveals distinct cytoskeletal architectures for cells invading through the HA matrix versus those migrating along the vascular channel. Substitution of collagen I in place of HA-RGD supports the same sequence of events but with faster local invasion and a more mesenchymal morphology. These results indicate that topographical effects are generalizable across matrix formulations, but that mechanisms underlying invasion are matrix-dependent. We anticipate that our reductionist paradigm should speed the development of mechanistic hypotheses that could be tested in more complex tumor models.

bioengineering↗