|
|
|
题名
|
作者
|
年代
|
出处
|
被引量
|
| 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 | Optimal cut-off values for obesity:using simple anthropometric indices to predict cardiovascular risk factors in Taiwan显示文摘 | Lee LT Chen CY | 2002 | Int J Obes Relat Metab Disord2002，26，： | 1 |
| 3 | Antioxidant and free radical scavenging effects of baicalein,baicalin and wogonin显示文摘 | Shieh DE Liu LT Lin CC | 2000 | Anticancer Res2000，20，5： | 1 |
| 4 | Simvastatin attenuates the additive effects of TNF-αand IL-18 on the connexin 43 up-regulation and over-proliferation of cultured aortic smooth muscle cells显示文摘 | Lin YC Chiang CH Chang LT | 2013 | Cytokine2013，62，3： | 1 |
| 5 | Yin Chen Hao Tang, a Chinese prescription, inhibits both herpes simplex virus type-1 and type-2 infections in vitro显示文摘 | Cheng HY Lin LT Huang HH | 2008 | Antiviral Res2008，77，1： | 1 |
| 6 | Melatonin preventshypertension and increased asymmetric dimethylargininein young spontaneous hypertensive rats显示文摘 | Tain YL Huang LT Lin IC | 2010 | J Pineal Res2010，49，4： | 1 |
| 7 | Pacific oyster-derived polysaccharides enhance antigen-specific T helper (Th)1 immunity in vitro and in vivo 显示文摘 | Cheng JY Ng LT Lin CL | 2013 | Immunopharmacol Immunotoxieol2013，35，2： | 1 |
| 8 | Prospective study of Helicabacter pylori eradication therapy in stage I(E) high-grade mucosa-associated lymphoid tissue lymphoma of the stomach显示文摘 | Chen LT Lin JT Shyu RY | 2001 | J Clin Oncol2001，19，22： | 1 |
| 9 | Preparation, physicochemical characterization, and antioxidant effects of quercetin nanoparticles显示文摘 | Wu TH Yen FL Lin LT | 2008 | International Journal of Pharmaceutics2008，346，12： | 1 |
| 10 | MDA5 plays a crucial role in enterovirus 71 RNA-mediated IRF3 activation 显示文摘 | Kuo RL Kao LT Lin SJ Wang RY Shih SR | 2013 | PLoS One2013，8，63： | 1 |
| 11 | Preparation, physicochemical charac- terization,and antioxidant effects of quercetin nanoparticles 显示文摘 | Wu TH Yen FL Lin LT | 2008 | Int J Pharm2008，346，12： | 1 |
| 12 | The correlation between insulin-like growth factorⅡmRNA binding protein 3 expression in hepatocellular carcinoma and prognosis显示文摘 | Chen LT Lin IJ Zheng LL | 2013 | Hepatogastroenterology2013，60，123： | 1 |
| 13 | Anti-melanogenic effects of 8-tocotrienol are associated with tyrosinase-related proteins and MAPK signaling pathway in B16 melanoma cells 显示文摘 | Ng LT Lin LT Chert CL | 2014 | Phytomedicine2014，21，3： | 1 |
| 14 | Effect of different extracts of Centella asiatica on cognition and markers of oxidative stress in rats显示文摘 | Liu LT Chiang LC | 2002 | J Ethnopharmacol2002，79，2： | 1 |
| 15 | The inhibitory effect of crystalline ricin in Ehrlich ascites tumor显示文摘 | Lin JY Lin LT Chen CC | 1970 | Taiwan Association J1970，69，1： | 1 |
| 16 | Antioxidant and free radical scavenging effects of bai-calein显示文摘 | Shieh DE Liu LT Lin CC | 2000 | Baicalin and wogonin Antic-ancer Res2000，20，5： | 1 |
| 17 | Optimal cut off values for obesi- ty: using simple anthropometric indices to predict cardiovascular risk factors in Taiwan显示文摘 | Lin WY Lee LT Chen CY | 2002 | lnt J Obes Relat Metab Disord2002，26，9： | 1 |
| 18 | Gene expression of cumulus cells in women with poor ovarian response after dehydroepiandrosterone supplementation显示文摘 | Tsui KH Lin LT Horng HC | 2014 | Taiwan J Obstet Gynecol2014，53，： | 1 |
| 19 | Metabolic syndrome is related to nonalcoholic steatohepatitis in severely obese subjects显示文摘 | Lin WY Lee LT | 2007 | Obes Surg2007，17，： | 1 |
| 20 | Optimal cut-off values for obesity:using simple anthropometric indices to predict cardiovascular risk factors in Taiwan显示文摘 | Lin WY Lee LT Chen CY | 2002 | Int J Obes Relat Metab Disord2002，26，9： | 1 |
您所在的机构暂未开通此部分的功能，请联系管理员