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Microfluidic isolation of highly pure embryonic stem cells using feeder-separated co-culture system.

ABSTRACT: 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.

SUBMITTER: Chen Q

PROVIDER: S-EPMC3743058 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Microfluidic isolation of highly pure embryonic stem cells using feeder-separated co-culture system.

Chen Qiushui Q Wu Jing J Zhuang Qichen Q Lin Xuexia X Zhang Jie J Lin Jin-Ming JM

Scientific reports 20130101

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 em ...[more]

PMID: 23942279

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Project description:Bioelectrochemical systems employing mixed microbial communities as biocatalysts are gaining importance as potential renewable energy, bioremediation, or biosensing devices. While we are beginning to understand how individual microorganism species interact with an electrode as electron donor, not much is known about the interactions between different microbial species in a community. Here, we compare the bioelectrochemical performance of Shewanella oneidensis in a pure-culture and in a co-culture with the homolactic acid fermenter Lactococcus lactis. While S. oneidensis alone can only use lactate as electron donor for current production, the co-culture is able to convert glucose into current with a similar coulombic efficiency of approximately 17%, respectively. With (electro)-chemical analysis and transcription profiling, we found that the BES performance and S. oneidensis physiology were not significantly different whether grown as a pure- or co-culture. These co-culture experiments represent a first step in understanding microbial interactions in BES communities with the goal to design complex microbial communities, which specifically convert target substrates into electricity. Further, for the first time, we elucidated S. oneidensis gene expression with an electrode as the only electron acceptor. The expression pattern confirms many previous studies regarding the enzymatic requirements for electrode respiration, and it generates new hypotheses on the functions of proteins, which are so far not known to be involved in electrode respiration. The BES was either operated with S. oneidensis alone, fed with lactate, or it was operated with S. oneidensis and L. lactis with glucose as primary substrate. The basic medium was a modified M4 medium containing 0.5 g/L yeast extract, 0.5 g/L trypton and 5 g/L glycerol phosphate, besides the commen M4 incredients. S. oneidensis oxidizes lactate to acetate and electrons in a BES - the latter generate a current at a graphite anode. The anode biofilm was harvested after about 4 weeks of continuous BES operation and subjected to total RNA extraction.

Dataset Information

Microfluidic isolation of highly pure embryonic stem cells using feeder-separated co-culture system.

Publications

Microfluidic isolation of highly pure embryonic stem cells using feeder-separated co-culture system.

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