Biology subjects

Jacob, A. G.

Publications and source records attributed to Jacob, A. G..

3 recordsLinked to original sources

bioRxiv2023

Differentiation and quality control of smooth muscle cells from human pluripotent stem cells via the neural crest lineage

The Sinha laboratory has developed protocols for differentiating human pluripotent stem cells (hPSCs) into vascular smooth muscle cells along developmental lineage-specific pathways. In development, paraxial mesoderm (PM), lateral plate mesoderm (LM) and neural crest (NC) linages each give rise to smooth muscle cells significant in a location-specific manner. Induced PSCs derived from patients enduring disease provide a platform from which disease-relevant cell models can be established in the laboratory. Here we describe a robust protocol for differentiating hPSCs into vascular smooth muscle cells via a neural crest lineage and the control steps required to ensure consistently high-quality differentiated cells.

cell biology↗

bioRxiv2021

Densified Collagen Tubular Grafts for Human Tissue Replacement and Disease Modelling Applications

There is a significant need across multiple indications for an off-the-shelf bioengineered tubular graft which fulfils the mechanical and biological requirements for implantation and function but does not necessarily require cells for manufacture or deployment. Herein, we present a tissue-like tubular construct using a cell-free, materials-based method of manufacture, utilizing densified collagen hydrogel. Our tubular grafts are seamless, mechanically strong, customizable in terms of lumen diameter and wall thickness, and display a uniform fibril density across the wall thickness and along the tube length. While the method enables acellular grafts to be generated rapidly, inexpensively, and to a wide range of specifications, the cell-compatible densification process also enables a high density of cells to be incorporated uniformly into the walls of the tubes, which we show can be maintained under perfusion culture. Additionally, the method enables tubes consisting of distinct cell domains with cellular configurations at the boundaries which may be useful for modelling aortic disease. Further, we demonstrate additional steps which allow for luminal surface patterning. These results highlight the universality of this approach and its potential for developing the next generation of bioengineered grafts.

bioengineering↗

bioRxiv2019

Identification of RBPMS as a smooth muscle master splicing regulator via proximity of its gene with super-enhancers

Alternative splicing (AS) programs are primarily controlled by regulatory RNA binding proteins (RBPs). It has been proposed that a small number of master splicing regulators might control cell-specific splicing networks and that these RBPs could be identified by proximity of their genes to transcriptional super-enhancers. Using this approach we identified RBPMS as a critical splicing regulator in differentiated vascular smooth muscle cells (SMCs). RBPMS is highly down-regulated during phenotypic switching of SMCs from a contractile to a motile and proliferative phenotype and is responsible for 20% of the AS changes during this transition. RBPMS directly regulates AS of numerous components of the actin cytoskeleton and focal adhesion machineries whose activity is critical for SMC function in both phenotypes. RBPMS also regulates splicing of other splicing, post-transcriptional and transcription regulators including the key SMC transcription factor Myocardin, thereby matching many of the criteria of a master regulator of AS in SMCs.

molecular biology↗