title: Microfluidic device capable of medium recirculation for non-adherent cell culture.
abstract: We present a microfluidic device designed for maintenance and culture of non-adherent mammalian cells, which enables both recirculation and refreshing of medium, as well as easy harvesting of cells from the device. We demonstrate fabrication of a novel microfluidic device utilizing Braille perfusion for peristaltic fluid flow to enable switching between recirculation and refresh flow modes. Utilizing fluid flow simulations and the human promyelocytic leukemia cell line, HL-60, non-adherent cells, we demonstrate the utility of this RECIR-REFRESH device. With computer simulations, we profiled fluid flow and concentration gradients of autocrine factors and found that the geometry of the cell culture well plays a key role in cell entrapping and retaining autocrine and soluble factors. We subjected HL-60 cells, in the device, to a treatment regimen of 1.25% dimethylsulfoxide, every other day, to provoke differentiation and measured subsequent expression of CD11b on day 2 and day 4 and tumor necrosis factor-alpha (TNF-α) on day 4. Our findings display perfusion sensitive CD11b expression, but not TNF-α build-up, by day 4 of culture, with a 1:1 ratio of recirculation to refresh flow yielding the greatest increase in CD11b levels. RECIR-REFRESH facilitates programmable levels of cell differentiation in a HL-60 non-adherent cell population and can be expanded to other types of non-adherent cells such as hematopoietic stem cells.
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Angela R
authors.initials: AR
authors.lastname: Dixon
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Shrinidhi
authors.initials: S
authors.lastname: Rajan
authors.affiliation: Department of Mechanical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA ; Mobility and Thermal Management Department, General Dynamics Land Systems, Sterling Heights, Michigan 48310, USA.
authors.forename: Chuan-Hsien
authors.initials: CH
authors.lastname: Kuo
authors.affiliation: Google, Inc., 1600 Amphitheatre Parkway Mountain View, California 94043, USA ; University of Michigan Comprehensive Cancer Center, Ann Arbor, Michigan 48109, USA.
authors.forename: Tom
authors.initials: T
authors.lastname: Bersano
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Wold
authors.affiliation: Department of Mechanical Engineering, Shibaura Institute of Technology, 3-5-1 Toyosu, Koto-ku, Tokyo 135-8548, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Futai
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA ; Department of Macromolecular Science and Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Takayama
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA ; Department of Materials Science and Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Geeta
authors.initials: G
authors.lastname: Mehta
references.citation: Blood. 1979 Sep;54(3):713-33
references.pmid: 288488
references.citation: Cell Mol Bioeng. 2010 Sep 1;3(3):213-228
references.pmid: 20824110
references.citation: Blood. 1990 Feb 1;75(3):626-32
references.pmid: 2153423
references.citation: Biomaterials. 2007 Jan;28(3):559-66
references.pmid: 16989897
references.citation: Lab Chip. 2006 Dec;6(12):1484-6
references.pmid: 17203151
references.citation: Anal Bioanal Chem. 2006 Jun;385(3):408-12
references.pmid: 16715270
references.citation: Biomaterials. 2008 Jun;29(17):2646-55
references.pmid: 18342367
references.citation: Lab Chip. 2007 Jun;7(6):756-62
references.pmid: 17538718
references.citation: Lab Chip. 2010 Apr 7;10(7):857-63
references.pmid: 20300672
references.citation: Anal Chem. 2009 May 15;81(10):3714-22
references.pmid: 19382754
references.citation: FEBS Lett. 1997 Aug 4;412(3):603-9
references.pmid: 9276475
references.citation: Ann Biomed Eng. 2008 Apr;36(4):554-62
references.pmid: 18172767
references.citation: Int Immunopharmacol. 2006 Jul;6(7):1204-13
references.pmid: 16714225
references.citation: Biomed Microdevices. 2007 Apr;9(2):123-34
references.pmid: 17160707
references.citation: Gene. 2008 Aug 1;419(1-2):16-26
references.pmid: 18547747
references.citation: Leuk Res. 1998 Jun;22(6):537-47
references.pmid: 9678720
references.citation: Methods Mol Biol. 2009;571:167-77
references.pmid: 19763966
references.citation: Integr Biol (Camb). 2009 Dec;1(11-12):649-54
references.pmid: 20027373
references.citation: Lab Chip. 2007 Nov;7(11):1497-503
references.pmid: 17960277
references.citation: Lab Chip. 2004 Oct;4(5):425-30
references.pmid: 15472725
references.citation: BMC Cell Biol. 2006 Feb 28;7:11
references.pmid: 16507101
references.citation: Anal Chim Acta. 2009 Sep 7;649(2):141-57
references.pmid: 19699390
references.citation: J Vasc Res. 2005 Jan-Feb;42(1):77-89
references.pmid: 15637443
references.citation: Lab Chip. 2006 Jan;6(1):149-54
references.pmid: 16372083
references.citation: Leukemia. 1992 Oct;6(10):1036-42
references.pmid: 1405756
references.citation: J Hematol Oncol. 2011 Apr 25;4:20
references.pmid: 21518431
references.citation: Biotechnol Bioeng. 2009 Aug 1;103(5):966-74
references.pmid: 19388086
references.citation: Biophys J. 2001 Oct;81(4):2020-34
references.pmid: 11566775
references.citation: BMC Syst Biol. 2009 Feb 18;3:20
references.pmid: 19222862
references.citation: Anal Chem. 1997 Apr 15;69(8):1564-8
references.pmid: 9109354
references.citation: J Leukoc Biol. 1985 Apr;37(4):407-22
references.pmid: 3855947
references.citation: Lab Chip. 2008 Oct;8(10):1700-12
references.pmid: 18813394
references.citation: Int J Hematol. 2008 Mar;87(2):189-194
references.pmid: 18256785
references.citation: Am J Physiol Endocrinol Metab. 2012 Feb 15;302(4):E389-95
references.pmid: 22114022
references.citation: J Orthop Res. 2009 Oct;27(10):1280-7
references.pmid: 19353691
references.citation: Annu Rev Anal Chem (Palo Alto Calif). 2008;1:423-49
references.pmid: 20636085
references.citation: Exp Hematol. 1987 Oct;15(9):928-35
references.pmid: 2820776
references.citation: Lab Chip. 2012 Jun 21;12(12):2240-6
references.pmid: 22402469
references.citation: Anal Chem. 2005 Jul 1;77(13):3993-9
references.pmid: 15987102
references.citation: Biotechnol Bioeng. 2006 Jun 20;94(3):596-609
references.pmid: 16586504
references.citation: Biophys J. 1995 Oct;69(4):1584-95
references.pmid: 8534829