Implementation of Plate Imaging for Demonstration of Monoclonality in Biologics Manufacturing Development.

Monoclonality of mammalian cell lines used for production of biologics is a regulatory expectation and one of the attributes assessed as part of a larger process to ensure consistent quality of the biologic. Historically, monoclonality has been demonstrated through statistics generated from limiting dilution cloning or through verified flow cytometry methods. A variety of new technologies are now on the market with the potential to offer more efficient and robust approaches to generating and documenting a clonal cell line.

A Strategy to Enhance Secretion of Extracellular Matrix Components by Stem Cells: Relevance to Tissue Engineering.

The ability of cells to secrete extracellular matrix proteins is an important property in the repair, replacement, and regeneration of living tissue. Cells that populate tissue-engineered constructs need to be able to emulate these functions. The motifs, KTTKS or palmitoyl-KTTKS (peptide amphiphile), have been shown to stimulate production of collagen and fibronectin in differentiated cells.

Assessment of Parylene C Thin Films for Heart Valve Tissue Engineering.

Background: Scaffolds are a key component of tissue-engineered heart valves (TEHVs). Several approaches had been adopted in the design of scaffolds using both natural and synthetic resources. We have investigated the suitability of parylene C (PC), a vapor deposited polymeric material, for the use as a scaffold in TEHV.

Shear stress and VEGF enhance endothelial differentiation of human adipose-derived stem cells.

Herein we combine chemical and mechanical stimulation to investigate the effects of vascular endothelial growth factor (VEGF) and physiological shear stress in promoting the differentiation human adipose derived stem cells (ADSCs) into endothelial cells. ADSCs were isolated and characterized; endothelial differentiation was promoted by culturing confluent cells in 50 ng/ml VEGF under physiological shear stress for up to 14 days. Afterwards, endothelial cells were seeded onto collagen or acellular aortic valve matrices and exposed to four culture conditions: shear stress + VEGF; shear stress - VEGF; static + VEGF and static - VEGF.

The potential of anisotropic matrices as substrate for heart valve engineering.

Cells environment is increasingly recognized as an important function regulator through cell-matrix interactions. Extracellular matrix (ECM) anisotropy being a key component of heart valves properties, we have devised a method to create highly porous anisotropic nanofibrillar scaffolds and studied their suitability as cell-support and interactions with human adipose derived stem cells (hADSCs) and human valve interstitial cells (hVICs). Anisotropic nanofibrillar scaffolds were produced by a modified jet-spraying method that allows the formation of aligned nanofibres (600 nm) through air-stream diffraction of a polymer solution (poly (ε-caprolactone, PCL) and collection onto a variably rotating drum.

MmNEU3 sialidase over-expression in C2C12 myoblasts delays differentiation and induces hypertrophic myotube formation.

Several factors affect the skeletal muscle differentiation process, in particular modifications of cell-cell contact, cell adhesion, and plasma membrane characteristics. In order to support the role of the plasma membrane-associated sialidase NEU3 in skeletal muscle differentiation and to analyse which events of the process are mainly affected by this sialidase, we decided to stably over-express MmNEU3 in C2C12 cells by a lentiviral vector and to investigate cell behavior during the differentiation process. Vitally stained C2C12 and NEU3 over-expressing cells were counted to reveal modifications in differentiation induction.

LPS-induced cytokine production in human dendritic cells is regulated by sialidase activity.

Removal of sialic acid from glycoconjugates on the surface of monocytes enhances their response to bacterial LPS. We tested the hypothesis that endogenous sialidase activity creates a permissive state for LPS-induced cytokine production in human monocyte-derived DCs. Of the four genetically distinct sialidases (Neu1-4), Neu1, Neu3, and Neu4 are expressed in human monocytes, but only Neu1 and Neu3 are up-regulated as cells differentiate into DCs.

Sialidase expression in activated human T lymphocytes influences production of IFN-gamma.

Sialidases influence cellular activity by removing terminal sialic acid from glycoproteins and glycolipids. Four genetically distinct sialidases (Neu1-4) have been identified in mammalian cells. In this study, we demonstrate that only lysosomal Neu1 and plasma membrane-associated Neu3 are detected in freshly isolated and activated human T lymphocytes.