Enhancement of insulin-producing cell differentiation from embryonic stem cells using pax4-nucleofection method

Abstract

Enhancement of IPC differentiation from EB-dissociated ES cells can be revealed by simply using pax4 expressing plasmid delivery. Not only more IPCs but also pancreatic differentiation-related genes can be detected by SQ-PCR. Expression of relative genes, such as foxa 2, mixl 1, pdx-1, insulin 1 and somatostatin after nucleofection, suggests that pax4 accelerates the whole differentiation progress. The higher insulin production with glucose dependent modulation suggests that pax4 expression can drive more mature IPCs. Although further determination of the entire mechanism is required, the potential of pax-4-nucleofected cells in medical treatment is promising.

Induction of Insulin-Producing Cells Derived from Endometrial Mesenchymal Stem-like Cells

“…EMSCs of passage 10 were used for phenotypic marker identification by flow cytometry (Lin et al, 2007). A total of 10 5 cells were resuspended in 100 l of PBS and incubated with primary antibodies (anti-human CD9, CD13, CD14, CD29, CD31, CD34, CD44, CD45, CD49, CD54, CD73, CD90, CD117, CD133, and STRO-1) (1:100 dilutions) at 4°C for 1 h. After washing twice with PBS, labeled cells were resuspended in 100 l of PBS with 1 l of the fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG antibody (AP124F; Millipore Corporation) at 4°C for 1 h. Cells were then examined with a FACSCalibur apparatus (BD Biosciences) (Lin et al, 2007).…”

“…Adult stem cells have yielded controversial results with regard to their ability to secrete insulin in vitro and normalize hyperglycemia in vivo. Several in vitro studies have shown that insulinproducing cells can be generated from adult pancreatic ductal tissues (Noguchi et al, 2005;Lin et al, 2006Lin et al, , 2007. Bone marrow-derived mesenchymal stem cells (BMSCs), which possess pluripotent differentiation capabilities, are a candidate for stem cell therapy in diabetic islet cell replacement .…”

Stem Cell Therapies for Type I Diabetes

“…When Pax4-transduced ESCs differentiated into IPCs and transplanted to diabetic mice, blood glucose levels were reduced for at least 4 weeks. Similar reduced blood glucose levels were detected in diabetic mice transplanted with IPCs transfected with Pax4 by nucleofection (Lin, Kao et al 2007). However, mice treated eithe r w i t h u n m o d i f i e d o r Pax4-modified ESC-IPCs developed teratomas (Blyszczuk, Asbrand et al 2004).…”

Effects of Intrahepatic Bone-Derived Mesenchymal Stem Cells Autotransplantation on the Diabetic Beagle Dogs

“…The regulation of insulin secretion by b cells is a complicated process of multi-signaling pathways involving many genes, including PDX-1, NEUROD-1, NGN-3, PAX-4, glucose transporter-2 (GLUT2), glucokinase, ATP-sensitive potassium channel, and the specific mechanism is not entirely clear at present [33][34][35][36][37][38][39]. Using genetic engineering technology, some researchers recently transferred PDX-1 and PAX gene into BMSCs, embryonic stem cells, and liver cells, and acquired the cells that were responsive to glucose stress in vitro [40][41][42]. However, these acquired insulinsecreting cells are still not comparable to mature b-cells due to their insufficiency of regulatory functions.…”