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| 1 | Simplified three-dimensional culture system for long-term expansion of embryonic stem cells显示文摘AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow fluorescent protein were cultured in three-dimensional(3-D) self-assembling scaffolds and compared with traditional two-dimentional(2-D) culture techniques requiring mouse embryonic fibroblast feeder layers or leukemia inhibitory factor. 3-D scaffolds encapsulating ESCs were prepared by mixing ESCs with polyethylene glycol tetra-acrylate(PEG-4-Acr) and thiolfunctionalized dextran(Dex-SH). Distribution of ESCs in 3-D was monitored by confocal microscopy. Viability and proliferation of encapsulated cells during long-term culture were determined by propidium iodide as well as direct cell counts and PrestoB lue(PB) assays. Genetic expression of pluripotency markers(Oct4, Nanog, Klf4, and Sox2) in ESCs grown under 2-D and 3-D cultureconditions was examined by quantitative real-time polymerase chain reaction. Protein expression of selected stemness markers was determined by two different methods, immunofluorescence staining(Oct4 and Nanog) and western blot analysis(Oct4, Nanog, and Klf4). Pluripotency of 3-D scaffold grown ESCs was analyzed by in vivo teratoma assay and in vitro differentiation via embryoid bodies into cells of all three germ layers. RESULTS: Self-assembling scaffolds encapsulating ESCs for 3-D culture without the loss of cell viability were prepared by mixing PEG-4-Acr and Dex-SH(1:1 v/v) to a final concentration of 5%(w/v). Scaffold integrity was dependent on the degree of thiol substitution of Dex-SH and cell concentration. Scaffolds prepared using Dex-SH with 7.5% and 33% thiol substitution and incubated in culture medium maintained their integrity for 11 and 13 d without cells and 22 ± 5 d and 37 ± 5 d with cells, respectively. ESCs formed compact colonies, which progressively increased in size over time due to cell proliferation as determined by confocal microscopy and PB staining. 3-D scaffold cultured ESCs expressed significantly higher levels(P < 0.01) of Oct4, Nanog, and Kl4, showing a 2.8, 3.0 and 1.8 fold increase, respectively, in comparison to 2-D grown cells. A similar increase in the protein expression levels of Oct4, Nanog, and Klf4 was observed in 3-D grown ESCs. However, when 3-D cultured ESCs were subsequently passaged in 2-D culture conditions, the level of these pluripotent markers was reduced to normal levels. 3-D grown ESCs produced teratomas and yielded cells of all three germ layers, expressing brachyury(mesoderm), NCAM(ectoderm), and GATA4(endoderm) markers. Furthermore, these cells differentiated into osteogenic, chondrogenic, myogenic, and neural lineages expressing Col1, Col2, Myog, and Nestin, respectively. CONCLUSION: This novel 3-D culture system demonstrated long-term maintenance of mouse ESCs without the routine passaging and manipulation necessary for traditional 2-D cell propagation. | Christina McKee Mick Perez-Cruet Ferman Chavez G Rasul Chaudhry | 2015 | World Journal of Stem Cells2015，7，7： | 2 |
| 2 | Performance of nanofihration membrane in a vibrating module (VSEP-NF) for arsenic removal 显示文摘 | Saber Ahmed Rasul M G | 2010 | Desalination2010，252，13： | 1 |
| 3 | Sustainability analysis of ecological and conventional agricultural systems in Bangladesh 显示文摘 | RASUL G THAPA G | 2003 | World Development2003，31，6： | 1 |
| 4 | Su- peracid catalyzed reactions of 5-am!no-l-naphthol with benzene and cyclohexane 显示文摘 | Koltunov K Y Surya Prakash G K Rasul G | 2002 | Tetrahedron2002，58，27： | 1 |
| 5 | Flow simulation in an electrostatic precipitator of a thermal power plant显示文摘 | Haque S M E Rasul M G Deev A V | 2009 | Applied Thermal Engineering2009，29，10： | 1 |
| 6 | Energy conservation measures in an institutional building in sub-tropical climate in Australia 显示文摘 | RAHMAN M M RASUL M G KHAN M M K | 2010 | Applied Energy2010，87，10： | 1 |
| 7 | Advances in heterogeneous photocatalytic degradation of phenols and dyes in wastewater 显示文摘 | Ahmed S Rasul M G Martens W N | 2011 | Water Air Soil Pollut2011，215，14： | 1 |
| 8 | Modeling and analysis of blast furnace performance for effi cient utilization of energy显示文摘 | Rasul M G Tanty B S Mohanty B | 2007 | Appl Therm Eng2007，27，1： | 1 |
| 9 | Sustainability Analysis of Ecological and Conventional Agricultural Systems in Bangladesh显示文摘 | Rasul G Thapa G | 2003 | World Development2003，31，6： | 1 |
| 10 | Flow simu- lation in an electrostatic precipitator of a thermal power plant 显示文摘 | Shah M E Haque Rasul M G Deev A V etal | 2009 | Applied Thermal Engineering2009，29，1： | 1 |
| 11 | Characterization of Azospirillum and relate diazotrophs associated with root of plant growing in saline soils显示文摘 | Bilal R Rasul G Qurshi J A | 1990 | World J of Microbiology and Biotechnology1990，11，6： | 1 |
| 12 | Alternative fuels for use in ce- ment kilns: Process impact modeling 显示文摘 | Kaddatz K T Rasul M G Rahman A | 2013 | Procedia Engineering2013，56，： | 1 |
| 13 | Influence of the inlet velocity profiles on the prediction of velocity distribution inside an electrostatic precipitator显示文摘 | Haque S M E Rasul M G Khan M M K | 2009 | Experimental Thermal and Fluid Science2009，33，2： | 1 |
| 14 | Flow simulation in an electrostatic precipitator of a thermal power plant显示文摘 | Shah M E Haque M G Rasul A V | 2009 | Applied Thermal Engineering2009，29，10： | 1 |
| 15 | Sustainability analysis of ecological and conventional agricultural systems in Bangladesh显示文摘 | Rasul G Thapa G | 2003 | World Development2003，31，6： | 1 |
| 16 | Implications of changing national policies on land use in the Chittagong Hill Tracts of Bangladesh显示文摘 | THAPA G B RASUL G | 2006 | Journal of Envi- ronmental Management2006，81，4： | 1 |
| 17 | Flow simulation in an electrostatic precipitator of a thermal power plant 显示文摘 | HAQUE S M E RASUL M G DEEV A V KHAN M M K | 2009 | Applied Thermal Engineering2009，29，10： | 1 |
| 18 | Effect of temperature rise on crop growth and productivi- ty显示文摘 | RASUL G CHAUDHRY Q Z MAHMOOD A | 2011 | Pakistan J Meteorol2011，8，15： | 1 |
| 19 | Beneficial effects of inoculated nitrogen-fixing bacteria on rice 显示文摘 | Mirz M S Rasul G Mehnaz S | 2000 | Biology and Fertility of Soils2000，31，： | 1 |
| 20 | Physical properties of bagasse显示文摘 | Rasul M G Rudolph V Carsky M | 1999 | Fuel1999，78，8： | 1 |
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