The present application provides materials and methods for treating a patient with one or more conditions or disorders associated with LAMA2, both ex vivo or in vivo. For example, the present disclosure provides materials and methods for treating a patient with Merosin-deficient Cogenital Muscular Dystrophy (MDCMD). The present application also provides materials and methods for editing a LAMA2 gene in a cell by genome editing. The present application also provides materials and methods for altering a contiguous genomic sequence of a LAMA2 gene in a cell. In addition, the present application provides one or more gRNAs for editing a LAMA2 gene. The present application also provides a therapeutic comprising at least one or more gRNAs for editing a LAMA2 gene. In addition, the present application provides a therapeutic for treating a patient with a LAMA2 related condition or disorder.

Described herein are cells, cell culture methods, and cell culture media compositions useful for producing and maintaining iPSC-derived cell lines that are of higher purity and maintain cell type integrity better than current iPSC-derived cell lines. Also disclosed are methods of using the described cells and media, such as therapeutic methods of use for the described cells. The described cells include iPSC-derived mesodermal precursor cells (MPC), which itself may differentiate into at least four different cell types. When cultured under appropriate conditions, the mesodermal precursor cells can be used to produce hematopoietic stem cells (HSC), mesenchymal stem cells (MSC), smooth muscle cells (SMC), or unlimited functional endothelial cells (UFEC). One characteristic that makes the described cells desirable is that they can be maintained in culture for a number of days, or passages, without changing phenotype through differentiation.

Co-culture of isolated human induced pluripotent stem cell (iPSC)-derived endothelial and smooth-muscle progenitor cells results in the formation of cellular spheroids having capillary-like structures (referred to herein as capillary fragments) at their core. Bioprinting of these spheroids into scaffold-free tissue constructs facilitates the development of microvasculature within the engineered tissue. Methods of using these bioprinted engineered tissues for cell therapy are also disclosed.

A method is disclosed for coating and patterning hydrogels in order to modify surface properties. The method exploits the water content of the hydrogel and the hydrophobicity of the reaction solvent to create a thin oxide adhesion layer on the hydrogel surface. This oxide adhesion layer enables rapid transformation of the hydrophilic, cell non-adhesive hydrogel into either a highly hydrophobic or a cell-adhesive hydrogel by reaction with an alkylphosphonic acid or an ,-diphosphonoalkane, respectively. Also disclosed are coated, patterned hydrogels and constructs comprising the coated, patterned hydrogels.

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105, CD90, CD66A, CD66E, CXCR4, CD133 or an SSEA. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1, TWIST, KLF-4 and Stella but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105, CD90, CD66A, CD66E, CXCF4, CD133 or an SSEA. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiating the stem cells, are also provided.

This invention relates, e.g., to a Myocyte-based Flow Assist Device (MFAD) for treating a subject in need of increased flow of a biological fluid, such as venous blood or lymph, comprising a sheath which comprises rhythmically contracting myocytes.

The invention relates to a hollow cell microfibre comprising successively, organized around a lumen, at least one endothelial cell layer, at least one smooth muscle cell layer, an extracellular matrix layer, and optionally an outer hydrogel layer. The invention also relates to a process for fabricating such a hollow cell microfibre.

CRISPR/Cas9-mediated genome editing holds clinical potential for treating genetic diseases, such as Duchenne muscular dystrophy (DMD), which is caused by mutations in the dystrophin gene. Here, using three promoters to drive expression of the same DMD guide RNA, a more robust and safe form of genome editing was achieved in a humanized mouse model for DMD with a deletion 13 in exon 50, and in a Ex50-MD Dog.

Provided are methods of isolating and culturing various cells in a living state including peripheral blood mononuclear cells (PBMCs) isolated from peripheral blood, which use a cell membrane permeabilization method. The methods use a streptococcal hemolytic exotoxin, which binds to cholesterol present in a cell membrane so as to make a pore therein, thereby allowing an exogenous protein to be permeated into the cell, and is a technique of isolating and culturing desired cells in a living state by probing a specific intracellular protein and performing flow cytometry (FACS). According to the methods, various intracellular proteins that could not previously be isolated and cultured from a patient's blood as well as various tissues may be targeted, and homogeneous cells may be obtained with high purity and high efficiency. Therefore, it is expected that the methods will highly contribute to many applications targeting a specific intracellular protein and research on mechanisms of various diseases and treatment thereof.

A multilayer scaffold device that includes a luminal electrospun layer, the luminal electrospun layer configured to provide a suitable environment to induce epithelium formation on the scaffold, an exterior electrospun layer, the exterior electrospun layer located radially exterior to the luminal electrospun layer, the exterior electrospun layer configured to induce formation of non-epithelial tissue; and at least one intermediate layer interposed between the luminal electrospun layer and that exterior electrospun layer, the intermediate layer configured to organize the formation of the respective epithelial tissue and the non-epithelial tissue.