An integrated cell culture lab on a chip: modular microdevices for cultivation of mammalian cells and delivery into microfluidic microdroplets.
We present a modular system of microfluidic PDMS devices designed to incorporate the steps necessary for cell biological assays based on mammalian tissue culture 'on-chip'. The methods described herein include the on-chip immobilization and culturing of cells as well as their manipulation by transfection. Assessment of cell viability by flow cytrometry suggests low attrition rates (<3%) and excellent growth properties in the device for up to 7 days for CHO-K1 cells. To demonstrate that key procedures from the repertoire of cell biology are possible in this format, transfection of a reporter gene (encoding green fluorescent protein) was carried out. The modular design enables efficient detachment and recollection of cells and allows assessment of the success of transfection achieved on-chip. The transfection levels (20%) are comparable to standard large scale procedures and more than 500 cells could be transfected. Finally, cells are transferred into microfluidic microdoplets, where in principle a wide range of subsequent assays can be carried out at the single cell level in droplet compartments. The procedures developed for this modular device layout further demonstrate that commonly used methods in cell biology involving mammalian cells can be reliably scaled down to allow single cell investigations in picolitre volumes.
Hufnagel
Hansjörg
H
Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, UK CB2 1GA.
Huebner
Ansgar
A
Gülch
Carina
C
Güse
Katharina
K
Abell
Chris
C
Hollfelder
Florian
F
D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation*; D049109:Cell Proliferation; D002470:Cell Survival; D018914:Cells, Immobilized / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D004867:Equipment Design; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D055616:Microtechnology / Q000295:instrumentation*; D014162:Transfection
D049452:Green Fluorescent Proteins