title: Reversible lineage-specific priming of human embryonic stem cells can be exploited to optimize the yield of differentiated cells.
abstract: The clinical use of human embryonic stem cells (hESCs) requires efficient cellular expansion that must be paired with an ability to generate specialized progeny through differentiation. Self-renewal and differentiation are deemed inherent hallmarks of hESCs and a growing body of evidence suggests that initial culture conditions dictate these two aspects of hESC behavior. Here, we reveal that defined culture conditions using commercial mTeSR1 media augment the expansion of hESCs and enhance their capacity for neural differentiation at the expense of hematopoietic lineage competency without affecting pluripotency. This culture-induced modification was shown to be reversible, as culture in mouse embryonic fibroblast-conditioned media (MEF-CM) in subsequent passages allowed mTeSR1-expanded hESCs to re-establish hematopoietic differentiation potential. Optimal yield of hematopoietic cells can be achieved by expansion in mTeSR1 followed by a recovery period in MEF-CM. Furthermore, the lineage propensity to hematopoietic and neural cell types could be predicted via analysis of surrogate markers expressed by hESCs cultured in mTeSR1 versus MEF-CM, thereby circumventing laborious in vitro differentiation assays. Our study reveals that hESCs exist in a range of functional states and balance expansion with differentiation potential, which can be modulated by culture conditions in a predictive and quantitative manner.
authors: McMaster Stem Cell and Cancer Research Institute, Michael G. DeGroote School of Medicine, McMaster University, Hamilton, Ontario, Canada.
authors: Jung Bok
authors: JB
authors: Lee
authors: Monica
authors: M
authors: Graham
authors: Tony J
authors: TJ
authors: Collins
authors: Jong-Hee
authors: JH
authors: Lee
authors: Seok-Ho
authors: SH
authors: Hong
authors: Amie Jamie
authors: AJ
authors: Mcnicol
authors: Zoya
authors: Z
authors: Shapovalova
authors: Mickie
authors: M
authors: Bhatia
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology; D000066449:Human Embryonic Stem Cells / Q000502:physiology*; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID
keywords: Developmental biology; Differentiation; Embryonic stem cells; Pluripotent stem cells; Self-renewal
references: Nat Biotechnol. 2002 Sep;20(9):933-6
references: 12161760
references: Nat Biotechnol. 2001 Oct;19(10):971-4
references: 11581665
references: Immunity. 2004 Jul;21(1):31-41
references: 15345218
references: Science. 1998 Nov 6;282(5391):1145-7
references: 9804556
references: Biol Reprod. 2005 Jan;72(1):42-9
references: 15317691
references: Blood. 2005 Jun 15;105(12):4598-603
references: 15718421
references: Biotechnol Bioeng. 2005 Sep 20;91(6):688-98
references: 15971228
references: Exp Hematol. 2005 Sep;33(9):987-96
references: 16140146
references: Hum Mol Genet. 2006 Jan 15;15(2):201-11
references: 16377660
references: Nat Biotechnol. 2006 Feb;24(2):185-7
references: 16388305
references: FEBS Lett. 2006 May 22;580(12):2875-8
references: 16716780
references: Nat Methods. 2006 Aug;3(8):637-46
references: 16862139
references: Methods Mol Biol. 2006;331:179-200
references: 16881518
references: Stem Cells. 2006 Oct;24(10):2170-6
references: 16741223
references: Hum Reprod. 2007 May;22(5):1231-8
references: 17251355
references: Nature. 2007 Aug 30;448(7157):1015-21
references: 17625568
references: Cell. 2007 Nov 30;131(5):861-72
references: 18035408
references: Science. 2007 Dec 21;318(5858):1917-20
references: 18029452
references: Nature. 2008 Jan 10;451(7175):141-6
references: 18157115
references: J Mol Med (Berl). 2008 Aug;86(8):875-86
references: 18521556
references: Cell Stem Cell. 2007 Nov;1(5):490-4
references: 18938745
references: Nat Biotechnol. 2009 Jan;27(1):91-7
references: 19122652
references: Stem Cell Res. 2010 May;4(3):165-79
references: 20363202
references: Cancer Res. 2010 Jun 1;70(11):4569-79
references: 20460541
references: Stem Cells Dev. 2010 Jun;19(6):753-61
references: 19686051
references: BMC Dev Biol. 2010;10:60
references: 20525219
references: Cell Reprogram. 2010 Apr;12(2):133-40
references: 20677928
references: Blood. 2011 Feb 3;117(5):1761-9
references: 21119109
references: Cell. 2011 Feb 4;144(3):439-52
references: 21295703
references: Nat Methods. 2011 May;8(5):424-9
references: 21478862
references: Cell Stem Cell. 2011 Jul 8;9(1):24-36
references: 21726831
references: Lancet. 2012 Feb 25;379(9817):713-20
references: 22281388
references: Stem Cells Dev. 2012 Oct 10;21(15):2866-77
references: 22548442
references: PLoS One. 2012;7(10):e48659
references: 23119085
references: Blood. 2003 Aug 1;102(3):906-15
references: 12702499