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 An Efficient Reprogramming Method for Human Induced Pluripotent Stem Cells (hiPSCs) Generation

 

  • Principal Investigator
    • Sung Ho Yoon, Biochemicals and Synthetic Biology Research Center / August 2013

  • R&D Background 
    •  High-throughput growth phenotyping is receiving great attention for establishing the genotype-phenotype map of sequenced organisms owing to the ready availability of complete genome sequences. To date, microbial growth phenotypes have been investigated mostly by the conventional method of batch cultivation using test tubes, Erlenmeyer flasks, or the recently available microwell plates. However, the current batch cultivation methods are time- and labor-intensive, and often fail to consider sophisticated environmental changes. The implementation of batch cultures at the nanoliter scale has been difficult because of the quick evaporation of the culture medium inside the reactors.

  • Research Summary 
    •  We devised a multiplexed microfluidic batch culture chip that enables microbial growth in 24 sets of discrete bioreactors (1 nL each), while simultaneously allowing the assessment of eight different culture conditions in parallel. The flow channels and culture reactors were designed to maintain a uniform distribution of cells in each reactor, and to conduct experiments in triplicates in a single run. The evaporation of the cell culture medium present in each of the batch reactors was overcome by placing the chip in a humidified incubator and incorporating anti-evaporation channels around the reactor within the chip. 

  • Applied Cases and Effects
    •  We demonstrate the versatility of the device by performing growth curve experiments with Escherichia coli and microbiological assays of antibiotics against the opportunistic pathogen Pseudomonas aeruginosa. Our study highlights that the microfluidic system can effectively replace the traditional batch culture methods with nanoliter volumes of bacterial cultivations, and it may be therefore promising for high-throughput growth phenotyping as well as for single-cell analyses.



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