Generation of renewable mouse intestinal epithelial cell monolayers and organoids for functional analyses.

Background: Conditional reprogramming has enabled the development of long-lived, normal epithelial cell lines from mice and humans by in vitro culture with ROCK inhibitor on a feeder layer. We applied this technology to mouse small intestine to create 2D mouse intestinal epithelial monolayers (IEC monolayers) from genetic mouse models for functional analysis.

Results: IEC monolayers form epithelial colonies that proliferate on a feeder cell layer and are able to maintain their genotype over long-term passage.

Aging effects on intestinal homeostasis associated with expansion and dysfunction of intestinal epithelial stem cells.

Intestinal epithelial stem cells (IESCs) are critical to maintain intestinal epithelial function and homeostasis. We tested the hypothesis that aging promotes IESC dysfunction using old (18-22 months) and young (2-4 month) Sox9-EGFP IESC reporter mice. Different levels of Sox9-EGFP permit analyses of active IESC (Sox9-EGFP), activatable reserve IESC and enteroendocrine cells (Sox9-EGFP), Sox9-EGFP progenitors, and Sox9-EGFP differentiated lineages.

Dystrophin-deficient cardiomyocytes derived from human urine: new biologic reagents for drug discovery.

The ability to extract somatic cells from a patient and reprogram them to pluripotency opens up new possibilities for personalized medicine. Induced pluripotent stem cells (iPSCs) have been employed to generate beating cardiomyocytes from a patient's skin or blood cells. Here, iPSC methods were used to generate cardiomyocytes starting from the urine of a patient with Duchenne muscular dystrophy (DMD).

Cloned, CD117 selected human amniotic fluid stem cells are capable of modulating the immune response.

Amniotic fluid stem (AFS) cells are broadly multipotent, can be expanded extensively in culture, are not tumorigenic and can be readily cryopreserved for cell banking. Mesenchymal stem cells (MSC) show immunomodulatory activity and secrete a wide spectrum of cytokines and chemokines that suppress inflammatory responses, block mixed lymphocyte reactions (MLR) and other immune reactions, and have proven therapeutic against conditions such as graft-versus-host disease. AFS cells resemble MSCs in many respects including surface marker expression and differentiation potential.

Inkjet-mediated gene transfection into living cells combined with targeted delivery.

In this study a novel method of simultaneous gene transfection and cell delivery based on inkjet printing technology is described. Plasmids encoding green fluorescent protein (GFP) were coprinted with living cells (porcine aortic endothelial [PAE] cells) through the ink cartridge nozzles of modified commercial inkjet printers. Agarose gel electrophoresis analysis showed there was no obvious structural alteration or damage to these plasmids after printing.