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| 1 | Enhancement of insulin-producing cell differentiation from embryonic stem cells using pax4-nucleofection method显示文摘AIM: To enhance the differentiation of insulin producing cell (IPC) ability from embryonic stem (ES) cells in vitro. METHODS: Four-day embryoid body (EB)-formatted ES cells were dissociated as single cells for the followed plasmid DNA delivery. The use of NucleofectorTM electroporator (Amaxa biosystems, Germany) in combination with medium-contained G418 provided a high efficiency of gene delivery for advanced selection. Neucleofected cells were plated on the top of fibronectincoated Petri dishes. Addition of Ly294002 and raised the glucose in medium at 24 h before examination.The differentiation status of these cells was monitored by semi-quantitative PCR (SQ-PCR) detection of the expression of relative genes, such as oct-4, sox-17, foxa2, mixl1, pdx-1, insulin 1, glucagons and somatostatin. The percentage of IPC population on d 18 of the experiment was investigated by immunohistochemistry (IHC), and the content/secretion of insulin was estimated by ELISA assay. The mice with severe combined immunodeficiency disease (SCID) pretreated with streptozotocin (STZ) were used to eliminate plasma glucose restoration after pax4+ ES implantation. RESULTS: A high efficiency of gene delivery was demonstrated when neucleofection was used in the present study; approximately 70% cells showed DsRed expression 2 d after neucleofection. By selection of medium-contained G418, the percentage of DsRed expressing cells kept high till the end of study. The pancreatic differentiation seemed to be accelerated by pax4 nucleofection. When compared to the group of cells with mock control, foxa2, mixl1, pdx1, higher insulin and somatostatin levels were detected by SQ-PCR 4 d after nucleofection in the group of pax4 expressing plasmid delivery. Approximately 55% of neucleofected cells showed insulin expression 18 d after neucleofection, and only 18% of cells showed insulin expression in mock control. The disturbance was shown by nucleofected pax4 RNAi vector; only 8% of cells expressed insulin 18 d after nucleofection. A higher IPC population was also detected in the insulin content by ELISA assay, and the glucose dependency was demonstrated in insulin secretion level. In the animal model, improvement of average plasma glucose concentration was observed in the group of pax-4 expressed ES of SCID mice pretreated with STZ, but no significant difference was observed in the group of STZ-pretreated SCID mice who were transplanted ES with mock plasmid. CONCLUSION: 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, thepotential of pax-4-nucleofected cells in medical treatment is promising. | Han-Tso Lin Chung-Lan Kao Kun-Hsiung Lee Yuh-Lih Chang Shih-Hwa Chiou Fu-Ting Tsai Tung-Hu Tsai Dey-Chyi Sheu Larry LT Ho Hung-Hai Ku | 2007 | World Journal of Gastroenterology2007，13，11： | 10 |
| 2 | Bioinpedance analysis of total body water in hemo-dialysis patients 显示文摘 | Ho LT Kushner RF Schoeller DA Gudivaka R SpiegelEM | 1994 | Kidney Int1994，46，： | 1 |
| 3 | Effect of green tea supplementation on insulin sensitivity in Sprague-Dawley rats显示文摘 | Wu LY Juan CC Ho LT | 2004 | Journal of Agricultural and Food Chemistry2004，52，3： | 1 |
| 4 | Selective suppression of insulin-induced proliferation of cultured human hepatoma cell by somatostatin显示文摘 | Chou CK Ho LT Ting LP | | 0，，： | 1 |
| 5 | Impact of ergocalciferol treatment of vitamin D deficiency on serum parathyroid hormone concentrations in chronic kidney disease 显示文摘 | Zisman AL Hristova M Ho LT | 2007 | Am J Nephrol2007，27，： | 1 |
| 6 | Rhabdomyolysis in diabetic emergencies 显示文摘 | Wang LM Tsai ST Ho LT | 1994 | Diabetes Res Clin Pract1994，26，3： | 1 |
| 7 | Neuroprotective effect of intermittent hypoxia on iron-induced oxidative injury in rat brain显示文摘 | Lin AM Chen CF Ho LT | 2002 | Exp Neurol2002，176，2： | 1 |
| 8 | The clinicopathological characteristics of gonadotroph cell adenoma: a study of 118 cases 显示文摘 | Ho DM Hsu CY Ting LT | 1997 | Hum Pathol1997，28，8： | 1 |
| 9 | Effect of green tea supplementation on insulin sensitivity in Sprague-Dawley rats显示文摘 | Wu LY Juan CC Ho LT | 2004 | Agric Food Chem2004，52，3： | 1 |
| 10 | Histopathology and MIB-1labeling index predicted recurrence of meningiomas:a proposal of diagnostic criteria for patients with aty pical meningioma显示文摘 | Ho DMT Hsu CY Ting LT | 2002 | Cancer2002，94，5： | 1 |
| 11 | Renal osteodystrophy in chronic renal failure显示文摘 | Ho LT Sprague SM | 2002 | Semin Nephrol2002，22，： | 1 |
| 12 | Effect of habitual physical activity on age-related glucose intolerance显示文摘 | Wang jT Ho LT | 1999 | J Am Geriatr Soc1999，37，： | 1 |
| 13 | Plurihormonal pituitary adenomas: immunostaining of all pituitary hormones is mandatory for correct classification 显示文摘 | Ho DM Hsu CY Ting LT | 2001 | Histopathology2001，39，2： | 1 |
| 14 | Effect of green tea supplementation on insulin sensitivity in Sprague-Dawley rats显示文摘 | Wu LY Juan CC Ho LT | 2004 | J Agile Food Chem2004，52，3： | 1 |
| 15 | Morbidity of cytomegalovirus infection in recipients of heart or heart-lung transplants who received cyclosporine显示文摘 | White LT Ho M | 1985 | J Infect Dis1985，152，6： | 1 |
| 16 | Promotion of cell affinity of porous PLLA scaffolds by immobilization of RGD peptides via plasma treatment 显示文摘 | Ho MH Hou LT Tu CY | 2006 | Macromol Biosci2006，1，： | 1 |
| 17 | Concurrent hyperthyroidism and hyperparathyroidism : influence of hyperthyroidism on serum magnesium, free calcium and parathyroid hormone显示文摘 | Lain HC Ho LT Tang KT etal | 1989 | Taiwan Yi Xue Hui Za Zhi1989，88，6： | 1 |
| 18 | Histopathology and MIB-1 labelling index predicted recurrence of meningiomas: aproposal of diagnostic criteria for patients with atypicalmeningioma显示文摘 | Ho DM Hsu CY Ting LT | 2002 | Cancer2002，94，5： | 1 |
| 19 | Histopathology and MIB-1 labeling index predicted recurrence of meningiomas: a proposal of diagnostic criteria for patients with atypical meningioma 显示文摘 | Ho DMT Hsu CY Ting LT | 2002 | Cancer2002，94，5： | 1 |
| 20 | Effects of exercise on insulin sensitivity, inflammatory cytokines, and serum tartrateresistant acid phosphatase 5a in obese Chinese male adolescents显示文摘 | Shih KC Janckila AJ Kwok CF Ho LT Chou YC Chao TY | 2010 | Metabolism2010，59，1： | 1 |
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