Pluripotent Cells : Culture

Pluripotent Cells Portfolios


Polymer Coating for Cell Culture Substrates

UW–Madison researchers have developed a new crosslinkable polymer coating for cell culture substrates. The nanometer-thin coating is made of glycidyl groups and azlactone groups distributed randomly along the copolymer backbone.

The coating is substrate independent and can be applied to a wide variety of organic and inorganic materials including plastic, silicon, glass and gold.

Patch for Delivering Therapeutic Cells into Heart

UW–Madison researchers have developed a new epicardial patch derived from cardiac-specific cells that can adhere to the heart without glue or sutures. The patch may be preloaded with stem cells, skeletal myoblasts or any other potentially therapeutic cell to treat injured or damaged hearts.

The patch is made of isolated three-dimensional cardiac extracellular matrix (ECM). Its composition mimics natural cardiac ECM. Along with other components, it is made up largely of the structural proteins fibronectin, collagen and elastin.

Peptide-Presenting Surfaces for Culturing Pluripotent Cells

UW–Madison researchers have developed a chemically defined, animal-free surface for culturing pluripotent cells. The surface is an insoluble substrate containing a peptide that binds to the cells and supports adhesion. More specifically, the peptide binds to a type of carbohydrate in the cells called glycosaminoglycan (GAG).

The researchers observed that pluripotent cells have surface receptors that recognize and adhere to GAG-binding peptides. They realized that the binding peptides could be displayed on a surface as a synthetic alternative to extracellular matrix proteins such as those in Matrigel®.

Furthermore, the new surfaces can be made to display combinations of adhesive ligands/epitopes with control over density, location and composition.

Clonal Culture of Human Pluripotent Stem Cells

UW–Madison researchers have developed a method to boost the cloning efficiency of any effective ES cell culture medium. The researchers screened small molecules and found they could increase efficiency rates using kinase enzyme inhibitors, e.g., protein kinase A/C/G inhibitors or a Rho-associated kinase (ROCK) inhibitor.

Microwells for Controlling Embryoid Body Formation

UW–Madison researchers have developed a three-dimensional microwell system that supports long-term embryonic stem cell culture and the formation of homogenous embryoid bodies. The microwells promote the growth of viable, undifferentiated embryonic stem cells that maintain pluripotency in culture for several weeks.

The use of micrometer-sized, dimensionally constrained wells helps control the size and shape of growing cell clumps, leading to more uniform EB formation. The exact dimensions of the wells can be varied so long as the shape and volume of the colonies cultured within the wells remain consistent.

Defined Surfaces of Self-Assembled Monolayers and Stem Cells

UW-Madison researchers have developed a method of identifying what peptides can be used to support a culture of hES cells. Self-assembled monolayer chemistry is used to create an array of alkane thiols on which peptide ligands attach in selected areas. Then cells, such as hES cells, are cultured with the monolayer as a substrate and the ligands that promote growth and self-renewal of cells are identifed.

Culturing Human Embryonic Stem Cells in Well Defined and Controlled Conditions

UW-Madison researchers have developed culture conditions for human ES cells that are entirely free of animal products, feeder cells and conditioned medium. These conditions permit the indefinite culture and proliferation of human ES cells in an undifferentiated state.

The medium includes high levels of a fibroblast growth factor, salts, vitamins, amino acids, glucose, gamma-aminobutyric acid, transforming growth factor beta, pipecholic acid, a lithium salt and lipids. It has been shown to support undifferentiated ES cell proliferation through at least thirteen passages and has proven sufficient to support the derivation of new lines of human ES cells.

The culture conditions also include a biological matrix in the culture vessel. The biological matrix is composed of four human proteins - collagen, laminin, fibronectin, and vitronectin.

No animal products of any kind and no fibroblasts from any species, including humans, are necessary in the culture. Instead, only purified, well characterized components are used to provide a more consistent culture system.