Cell biology is different in small volumes: endogenous signals shape phenotype of primary hepatocytes cultured in microfluidic channels.
The approaches for maintaining hepatocytes in vitro are aimed at recapitulating aspects of the native liver microenvironment through the use of co-cultures, surface coatings and 3D spheroids. This study highlights the effects of spatial confinement-a less studied component of the in vivo microenvironment. We demonstrate that hepatocytes cultured in low-volume microfluidic channels (microchambers) retain differentiated hepatic phenotype for 21 days whereas cells cultured in regular culture plates under identical conditions de-differentiate after 7 days. Careful consideration of nutrient delivery and oxygen tension suggested that these factors could not solely account for enhanced cell function in microchambers. Through a series of experiments involving microfluidic chambers of various heights and inhibition of key molecular pathways, we confirmed that phenotype of hepatocytes in small volumes was shaped by endogenous signals, both hepato-inductive growth factors (GFs) such as hepatocyte growth factor (HGF) and hepato-disruptive GFs such as transforming growth factor (TGF)-β1. Hepatocytes are not generally thought of as significant producers of GFs-this role is typically assigned to nonparenchymal cells of the liver. Our study demonstrates that, in an appropriate microenvironment, hepatocytes produce hepato-inductive and pro-fibrogenic signals at the levels sufficient to shape their phenotype and function.
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Amranul
A
Haque
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Pantea
P
Gheibi
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Yandong
Y
Gao
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Elena
E
Foster
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Kyung Jin
KJ
Son
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.|Department of Plant and Environmental New Resources, Kyung Hee University, Youngin-si, Gyeonggi-do, South Korea.
Jungmok
J
You
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Gulnaz
G
Stybayeva
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Dipali
D
Patel
Department of Biomedical Engineering, University of California Davis, CA 95616, USA.
Alexander
A
Revzin
Liver. 1999 Apr;19(2):151-9
10220746
Gene Ther. 2000 Feb;7(3):205-14
10694797
FASEB J. 1989 Feb;3(2):174-7
2914628
Cells Tissues Organs. 2005;181(2):67-79
16534201
Biomaterials. 2011 Mar;32(8):2032-42
21163520
Mol Pharmacol. 2002 Jul;62(1):65-74
12065756
Biomaterials. 2012 Feb;33(5):1414-27
22118818
Cell Res. 2009 Feb;19(2):156-72
19153598
Biomaterials. 2010 May;31(13):3596-603
20153045
J Biomed Mater Res A. 2005 Oct 1;75(1):242-8
16052491
Nature. 2014 Mar 13;507(7491):181-9
24622198
Hepatology. 2008 Jul;48(1):276-88
18536058
Am J Physiol Cell Physiol. 2011 Mar;300(3):C416-24
21160029
Lab Chip. 2014 Jun 21;14(12):2033-9
24770663
Cell Biol Toxicol. 1997 Jul;13(4-5):235-42
9298244
Organogenesis. 2014 Apr-Jun;10(2):250-9
25102189
Tissue Eng Part A. 2013 Dec;19(23-24):2655-63
23815179
Hepatology. 2001 Sep;34(3):447-55
11526528
Biotechnol Bioeng. 2013 Jun;110(6):1663-73
23280535
Biotechnol Bioeng. 2007 Aug 1;97(5):1340-6
17286266
Hepatology. 1996 Sep;24(3):636-42
8781336
Ann Surg Treat Res. 2014 Aug;87(2):53-60
25114883
Biotechnol Prog. 1991 May-Jun;7(3):237-45
1367596
Hepatology. 1984 May-Jun;4(3):373-80
6373549
Nat Rev Mol Cell Biol. 2014 Mar;15(3):178-96
24556840
Ann Surg. 1994 Jul;220(1):59-67
8024360
Lab Chip. 2005 Oct;5(10):1089-95
16175265
Science. 1997 Apr 4;276(5309):60-6
9082986
Biotechnol Bioeng. 2008 Apr 15;99(6):1472-81
17969156
Biomaterials. 1996 Feb;17(3):373-85
8745335
Biotechnol Bioeng. 2003 May 5;82(3):253-62
12599251
Biomaterials. 2009 Aug;30(22):3733-41
19375794
Nat Methods. 2005 Feb;2(2):119-25
15782209
J Gastroenterol Hepatol. 2008 Mar;23 Suppl 1:S122-7
18336655
J Clin Invest. 1995 Jul;96(1):447-55
7615817
Biomed Microdevices. 2011 Jun;13(3):539-48
21424383
Nature. 1970 Jan 31;225(5231):420-2
5411117
FASEB J. 1996 Nov;10(13):1471-84
8940293
Ann N Y Acad Sci. 1980;349:85-98
7013612
Biotechnol Bioeng. 2007 May 1;97(1):188-99
17054120
Hepatology. 1996 Nov;24(5):1274-81
8903410
J Biomed Mater Res. 1997 Feb;34(2):189-99
9029299
J Vis Exp. 2013 Jul 15;(77):e50678
23893025
ACS Appl Mater Interfaces. 2015 Jun 17;7(23):12299-308
25247391
Annu Rev Anal Chem (Palo Alto Calif). 2012;5:293-315
22524217
Integr Biol (Camb). 2014 Jan;6(1):44-52
24247788
Tissue Eng Part A. 2011 Apr;17(7-8):1055-68
21121876
FASEB J. 2006 Apr;20(6):773-5
16484330
Biotechnol Bioeng. 2005 Jun 5;90(5):632-44
15834948
J Biochem. 2011 Mar;149(3):231-9
21217146
Hepatology. 2011 Jun;53(6):2063-74
21384409
Hepatology. 2002 Aug;36(2):336-44
12143041
Anal Chem. 2006 Jul 1;78(13):4291-8
16808435
Lab Chip. 2005 Jan;5(1):44-8
15616739
Biomaterials. 2005 Jun;26(17):3607-15
15621251
Stem Cells. 2016 Jun;34(6):1501-12
26865369
J Cell Biol. 1973 Dec;59(3):722-34
4357460
Nat Biotechnol. 2008 Jan;26(1):120-6
18026090
Science. 2006 Oct 13;314(5797):298-300
17038622
PLoS One. 2011;6(8):e22892
21829665
Lab Chip. 2015 Dec 21;15(24):4614-24
26542093
Exp Mol Med. 2001 Dec 31;33(4):179-90
11795478
Expert Opin Investig Drugs. 1999 May;8(5):585-607
15992118
Biotechnol Bioeng. 1997 Dec 20;56(6):706-11
18642343
Annu Rev Cell Dev Biol. 2011;27:377-407
21801015
Integr Biol (Camb). 2009 Feb;1(2):182-95
20023802