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1、细胞图型及微流控芯片上的细胞培养和分化【摘要】：微流控和图型化技术在细胞生物学领域中的应用具有重要的意义。本论文首先讨论几种新的细胞图型化技术。其一为PDMS 模负压吸附法,由此形成的物理或化学印迹被用于胚胎纤维细胞的图型化;其二为高分辨率压印刻蚀法,通过紫外光软膜纳米压印技术可制作高分辨率生物分子图型,细胞的贴附情况表明此法有效地改善了生物分子图型的稳定性;另外微流控芯片中单细胞阵列也被实现,并且细胞可以长时间的培养观察。微流控技术首先被用于基底材料对胚胎干细胞的培养和成神经细胞分化影响的研究,结果表面,尽管胚胎干细胞可在多种基底上生长,细胞贴附力较强的基底更利于分化实验的实现。微流控芯片也
2、被用于研究骨髓间充质干细胞的培养及相应的成脂肪细胞和神经细胞的分化。且由微接触印刷法制备的图型被用于诱导骨髓间充质干细胞的成神经细胞分化。又制作了用于老鼠胚胎干细胞的神经分化的微井图型。最后,尝试了神经细胞电极芯片,用于信号传导的测量。 【关键词】：微流控纳米制作细胞培养和分化干细胞【学位授予单位】：华东师范大学【学位级别】：博士【学位授予年份】：2010【分类号】：Q813【目录】：Acknowledgement6-8Abstract8-9AbstractinFrench9-10 摘要 10-11Tableofcontents11-15Outlines15-17Chapter1Introdu
3、ction17-411.1CellPatterning17-261.1.1Cellsandstemcells17-201.1.2Cell-materialinteraction20-211.1.3Previousworks21-261.2MicrofluidicsandLab-on-achipsystem26-361.2.1HistoryrecallofMicrofludics26-301.2.2FundamentalnotionsofMicrofluidics30-321.2.3Onchipcellcultureanddifferentiation32-331.2.4Previouswork
4、s33-361.3Objectiveofthiswork36-37References37-41Chapter2Cellpatterning41-752.1Microfabricationtechniques41-492.1.1Photolithography41-442.1.2Softlithography44-452.1.3Microcontactprinting(CP)45-472.1.4Nanoimprintlithography(NIL)47-492.2PDMSdegasinducedcellpatterning49-562.2.1PDMSmoldfabrication512.2.2
5、PDMSdegasinducedpatterining51-522.2.3Cellpatterning52-532.2.4Resultsanddiscussions53-562.2.5Conclusions562.3Highresolutioncellpatterning56-672.3.1Generallithographymethod57-592.3.2Culturepatterning59-602.3.3Resultsanddiscussions60-662.3.4Conclusions66-672.4Singlecellpatterninmicrofluidicchip67-722.4
6、.1Thefabricationofthepatternedchip67-692.4.2ThePoly-Lysine-FITCpattern69-702.4.3Singlecellpatterninchip70-722.5Conclusion72-73References73-75Chapter3On-chipcellculture:easeofEScells75-893.1Introduction75-763.2MicrofluidicdevicefabricationformEScells76-773.3SimpledevicesforMEFs773.4On-chipESculturean
7、ddifferentiation77-783.4.1mESCellculture77-783.4.2EBformationanddifferentiation783.4.3SeedingMEFcellsonmicro-channel-wellsystem783.5Immunocytochemistry78-793.6SeedingMEFsonthedifferentratiosofPDMSsubstrates793.7ResultsanddiscussionsforEScells79-843.7.1Celladhesiondependsondifferentsubstrates79-823.7
8、.2ESco-culturewithMEFontomicro-channel-wellsystem82-833.7.3ES-derivedneuraldifferentiationonmicro-channel-wellsystem83-843.8ResultsanddiscussionsforMEFsonPDMSsubstrates84-873.9Conclusion87-88References88-89Chapter4On-chipcellculture:caseofMSCs89-1034.1Introduction89-904.2Microfluidicdevicefabricatio
9、n90-934.3On-chipcellculture93-944.4RMSCscultureinthethreemicrofluidicdevices94-1024.4.1CharacterizationofmicrofluidicdevicesforRMSCs94-954.4.2RMSCsinmicrofluidicdevice95-964.4.3RMSCsinmicrofluidicdevice96-974.4.4RMSCsinmicrofluidicdevice97-994.4.5RMSCsincultureflask99-1004.4.6Comparisonofthethreedev
10、ices100-102References102-103Chapter5On-chipcelldifferentiation:caseofMSCs103-1255.1Introduction103-1045.2Mierofluidicdevicefabrication104-1065.3Differentiationprotocol106-1095.3.1Adipogenicdifferentiation106-1075.3.2Neuraldifferentiation107-1085.3.3Characterizationtechniques108-1095.4Resultsanddiscu
11、ssions109-1215.4.1Adipogenicdifferentiation109-1135.4.2Neuraldifferentiation113-1215.5Conclusion121-122References122-125Chapter6Towardsacellularnetworkformation125-1416.1Introduction125-1266.2Experiments126-1296.2.1MicrocontactprintingpatternforRMSCs126-1286.2.2Themicrowell-networkformEScells128-129
12、6.3MicrocontactprintingPLLPatternforRMSCs129-1326.3.1MicrocontactprintingPLLPattern129-1306.3.2RMSCsnetworkpattern130-1326.3.3Otherpatterns1326.3.4Concludingremarks1326.4Mirowell-networkformEScells132-1366.4.1themicrowell-network132-1336.4.2ruES-derivedneuronsinthemicrowellnetwork133-1366.5Firsttryo
13、nelectrodechipforthesignaltransduction136-140References140-141SummaryandPerspective141-145Publicationlistofthisthesiswork145-147APPENDIX147-178AppendixPotocolofcultureanddifferentiationofRMSCs147-151AppendixPotocolofcultureanddifferentiationofhMSCs151-155AppendixProtocolofcultureanddifferentiationofmouseES155-161AppendixSelfloadingandcellculture161-169AppendixDifferentiationofhMSCstoosteoblastes169-178 本论文购买请联系页眉网站。
