Microfluidic isolation of highly pure embryonic stem cells using feeder-separated co-culture system.
Engineered artificial tissues from stem cells show great potential in regenerative medicine, disease therapies and organ transplantation. To date, stem cells are typically co-cultured with inactivated feeder layers to maintain their undifferentiated state, and to ensure reliable cell purity. Herein, we propose a novel microfabricated approach for feeder-separated coculture of mouse embryonic stem (mES) cells on polydimethylsiloxane (PDMS) porous membrane-assembled 3D-microdevice. Normal mouse embryonic fibroblasts (mEFs) without inactivation were specifically co-cultured with mES cells, resulting in the formation of mES cell colonies on spatially controlled co-culture with feeder layers. An excellent undifferentiated state was confirmed by the expressions of Nanog, octamer binding protein 4 (Oct-4) and alkaline phosphatase (ALP) after 5 days culture. As a result, with the significant advantages of efficiency and simplicity, pure mES cell populations (a purity of 89.2%) from mEFs co-cultures were easily collected without any further purification or separation.
Department of Chemistry, Beijing Key Laboratory Microanalysis and Instrumentation, Tsinghua University, Beijing 100084, P.R. China.
Qiushui
Q
Chen
Jing
J
Wu
Qichen
Q
Zhuang
Xuexia
X
Lin
Jie
J
Zhang
Jin-Ming
JM
Lin
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000201:enzymology; D004867:Equipment Design; D061252:Feeder Cells / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D055616:Microtechnology; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism
D004129:Dimethylpolysiloxanes; D050814:Octamer Transcription Factor-3; D000469:Alkaline Phosphatase