Jump to main content
Jump to site search
PLANNED MAINTENANCE Close the message box

Scheduled maintenance upgrade on Thursday 4th of May 2017 from 8.00am to 9.00am (BST).

During this time our websites will be offline temporarily. If you have any questions please use the feedback button on this page. We apologise for any inconvenience this might cause and thank you for your patience.


Issue 22, 2013
Previous Article Next Article

Optimal periodic perfusion strategy for robust long-term microfluidic cell culture

Author affiliations

Abstract

Long-term cell culture in microfluidic devices is an essential prerequisite for “on a chip” biological and physiological based studies. We investigated how medium delivery, from continuous to periodic perfusion, affects long-term cell cultures in a microfluidic platform. Computational simulations suggested that different delivery strategies result in different temporal profiles of accumulation and washing out of endogenous (EnF) and exogenous (ExF) factors, respectively. Thus, cultures exposed to the same overall amount of medium with different temporal profiles were analysed in terms of homogeneity, cell morphology and phenotype. Murine and human cell lines (C2C12 and HFF) and mouse embryonic stem cells (mESC) were cultured in microfluidic channels. An ad hoc experimental setup was developed to perform continuous and periodic medium delivery into the chip, tuning the flow rate, the perfusion time, and the interval of perfusion while using the same amount of medium volume. Periodic medium delivery with a short perfusion pulse ensured cell homogeneity compared to standard cell culture. Conversely, a continuous flow resulted in cell heterogeneity, with abnormal morphology and vesiculation. Only dramatic and unfeasible increasing of perfused medium volume in the continuous configuration could rescue normal cell behaviour. Consistent results were obtained for C2C12 and HFF. In order to extend these results to highly sensitive cells, mESC were cultured for 6 days in the microfluidic channels. Our analysis demonstrates that a periodic medium delivery with fast pulses (with a frequency of 4 times per day) resulted in a homogeneous cell culture in terms of cell viability, colony morphology and maintenance of pluripotency markers. According to experimental observations, the computational model provided a rational description of the perfusion strategies and of how they deeply shape the cell microenvironment in microfluidic cell cultures. These results provide new insight to define optimal strategies for homogeneous and robust long-term cell culture in microfluidic systems, an essential prerequisite for lab on chip cell-based applications.

Graphical abstract: Optimal periodic perfusion strategy for robust long-term microfluidic cell culture

Back to tab navigation
Please wait while Download options loads

Supplementary files

Publication details

The article was received on 27 May 2013, accepted on 22 Aug 2013 and first published on 22 Aug 2013


Article type: Paper
DOI: 10.1039/C3LC50643F
Citation: Lab Chip, 2013,13, 4430-4441
  •   Request permissions

    Optimal periodic perfusion strategy for robust long-term microfluidic cell culture

    S. Giulitti, E. Magrofuoco, L. Prevedello and N. Elvassore, Lab Chip, 2013, 13, 4430
    DOI: 10.1039/C3LC50643F

Search articles by author