Quantitative Analysis of Apoptosis and Necrosis in Live Cells Using Flow Cytometry.

Apoptosis and necrosis are the two sides of the cell death penumbra. Apoptosis is a well-studied model of cell death wherein the cell destroys itself employing a predefined form of active signaling without the release of soluble cytoplasmic contents to the external environment. Compared to apoptosis, necrosis is a nonspecific form of sudden cell death in response to an invasive external stimulus which in turn is devoid of active programmed intracellular signaling leading to the sudden release of the soluble cellular contents consequent to the rupture of the cell membrane.

Drug loaded microbeads entrapped electrospun mat for wound dressing application.

A new design of antibiotic loaded wound dressing and its initial in vitro evaluation is described. Chitosan microbeads loaded with ampicillin were sandwiched within polycaprolactone electrospun mat (MbAPPCL). The morphology was analyzed by scanning electron microscopy and surface chemistry was characterized by Fourier Transform Infrared Spectroscopy.

A quantitative real-time approach for discriminating apoptosis and necrosis.

Apoptosis and necrosis are the two major forms of cell death mechanisms. Both forms of cell death are involved in several physiological and pathological conditions and also in the elimination of cancer cells following successful chemotherapy. Large number of cellular and biochemical assays have evolved to determine apoptosis or necrosis for qualitative and quantitative purposes.

Characterization and in vitro evaluation of electrospun chitosan/polycaprolactone blend fibrous mat for skin tissue engineering.

The electrospinning technique allows engineering biomimetic scaffolds within micro to nanoscale range mimicking natural extracellular matrix (ECM). Chitosan (CS) and polycaprolactone (PCL) were dissolved in a modified solvent mixture consisting of formic acid and acetone (3:7) and mixed in different weight ratios to get chitosan-polycaprolactone [CS-PCL] blend solutions. The CS-PCL blend polymer was electrospun in the same solvent system and compared with PCL.

Identification of human corneal epithelial stem cells on the basis of high ABCG2 expression combined with a large N/C ratio.

Purpose: Till date there is no exclusive marker for human corneal epithelial stem cells (CESCs). In this study, our strategy is to combine high expression of ABCG2, a putative SC marker with high N/C ratio to develop a specific method for identification of CESCs.

Methods: Limbal/corneal epithelial cells (LECs/CECs) were isolated from cadaver eyes by enzymatic treatment and the cytospin smears/cryosections were immunostained for ABCG2, p63, or connexin-43 (Cx-43) and counterstained with propidium iodide/DAPI.

Intelligent thermoresponsive substrate from modified overhead projection sheet as a tool for construction and support of cell sheets in vitro.

Cell sheet engineering using thermoresponsive culture dishes allows harvesting of intact in vitro cell sheet. In this study, commercially available polyethylene terephthalate-based overhead projection transparency sheet (OHPS) was identified as a substrate for coating thermoresponsive poly(N-isopropylacrylamide-co-glycidylmethacrylate) (NGMA) copolymer having lower critical solution temperature of 28°C. Since OHPS is highly hydrophobic and rigid, the surface was modified by alkali treatment (OHPS-M) to functionalize the surface with carboxyl and hydroxyl groups so as to make it more suitable for efficient coating of NGMA copolymer and cell culture.

Optimization of culture conditions for an efficient xeno-feeder free limbal cell culture system towards ocular surface regeneration.

Ex vivo expansion of limbal stem cells from a small biopsy and its subsequent transplantation is the golden choice of treatment for limbal stem cell deficiency. Use of murine 3T3 feeder layer is a prerequisite for this ex vivo expansion. There is an ever-increasing demand for feeder free cultures to avoid xenotoxicity and transmission of xeno-diseases to human system.