Hypoxic Stress Forces Irreversible Differentiation of a Majority of Mouse Trophoblast Stem Cells Despite FGF4.
Hypoxic, hyperosmotic, and genotoxic stress slow mouse trophoblast stem cell (mTSC) proliferation, decrease potency/stemness, and increase differentiation. Previous reports suggest a period of reversibility in stress-induced mTSC differentiation. Here we show that hypoxic stress at 0.5% O2decreased potency factor protein by ∼60%-90% and reduced growth to nil. Hypoxia caused a 35-fold increase in apoptosis at Day 3 and a 2-fold increase at Day 6 above baseline. The baseline apoptosis rate was only 0.3%. Total protein was never less than baseline during hypoxic treatment, suggesting 0.5% O2is a robust, nonmorbid stressor. Hypoxic stress induced ∼50% of trophoblast giant cell (TGC) differentiation with a simultaneous 5- to 6-fold increase in the TGC product antiluteolytic prolactin family 3, subfamily d, member 1 (PRL3D1), despite the presence of fibroblast growth factor 4 (FGF4). Hypoxia-induced TGC differentiation was also supported by potency and differentiation mRNA marker analysis. FGF4 removal at 20% O2committed cell fate towards irreversible differentiation at 2 days, with similar TGC percentages after an additional 3 days of culture under potency conditions when FGF4 was readded or under differentiation conditions without FGF4. However, hypoxic stress required 4 days to irreversibly differentiate cells. Runted stem cell growth, forced differentiation of fewer cells, and irreversible differentiation limit total available stem cell population. Were mTSCs to respond to stress in a similar mode in vivo, miscarriage might occur as a result, which should be tested in the future.
CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan.|Department of Physiology, Wayne State University School of Medicine, Detroit, Michigan.
Yu
Y
Yang
CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan.|Perinatology Research Branch/NICHD/NIH, Wayne State University, Detroit, Michigan.|Department of Molecular Biomedicine, CINVESTAV-IPN, Mexico City, Mexico.
Marcia
M
Arenas-Hernandez
CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan.|Perinatology Research Branch/NICHD/NIH, Wayne State University, Detroit, Michigan.
Nardhy
N
Gomez-Lopez
CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan.
Jing
J
Dai
Department of Pediatrics and Children's Hospital of Michigan, Wayne State University School of Medicine, Detroit, Michigan.
Graham C
GC
Parker
CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan.
Elizabeth E
EE
Puscheck
CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan daniel.rappolee@med.wayne.edu.|Department of Physiology, Wayne State University School of Medicine, Detroit, Michigan.|Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan.|Institutes for Environmental Health Science, Wayne State University School of Medicine, Detroit, Michigan.|Department of Biology, University of Windsor, Windsor, Ontario, Canada.
Daniel A
DA
Rappolee
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D015687:Cell Hypoxia / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D051519:Fibroblast Growth Factor 4 / Q000378:metabolism*; D051379:Mice; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism*
D051519:Fibroblast Growth Factor 4
differentiation; hypoxic stress; potency; trophoblast stem cells
Hum Reprod Update. 2002 Jul-Aug;8(4):333-43
12206468
Mol Reprod Dev. 1993 Apr;34(4):357-68
8471259
Endocrinology. 1994 Jun;134(6):2390-6
8194465
BMC Genomics. 2008 Jul 28;9:352
18662396
J Cell Biol. 1999 Dec 27;147(7):1431-42
10613902
N Engl J Med. 1988 Jul 28;319(4):189-94
3393170
Dev Biol. 2011 Oct 1;358(1):231-9
21839735
Genes Dev. 2008 Nov 1;22(21):3024-36
18981479
Am J Public Health. 1997 Jun;87(6):1003-7
9224184
Recent Prog Horm Res. 2002;57:221-34
12017545
Placenta. 2009 Jan;30(1):66-73
19036436
Trends Biochem Sci. 2008 Mar;33(3):141-50
18291657
Reproduction. 2010 Dec;140(6):921-30
20876741
Endocrinology. 1991 Dec;129(6):2895-906
1954876
Hypertension. 2008 Apr;51(4):970-5
18259009
Placenta. 2011 Jun;32(6):475-81
21511332
Biol Reprod. 2016 Jun;94(6):122
27122635
Methods Mol Med. 2006;122:427-34
16511999
Biol Reprod. 2006 Jun;74(6):1041-50
16481593
Biochemistry. 1989 Mar 21;28(6):2631-8
2471551
Placenta. 2006 Aug;27(8):869-75
16289730
Stem Cell Res. 2014 Nov;13(3 Pt A):478-91
25239494
Dev Dyn. 2003 Jun;227(2):185-91
12761846
Acta Physiol Hung. 2009 Jun;96(2):243-50
19457768
J Biol Chem. 2005 Apr 29;280(17):16925-33
15684421
Development. 2000 Oct;127(19):4195-202
10976051
Development. 1995 Aug;121(8):2513-23
7671815
J Reprod Dev. 2015;61(4):325-31
25985793
Mol Biol Cell. 1998 Apr;9(4):795-807
9529378
Nat Genet. 1998 Mar;18(3):271-5
9500551
Development. 2000 Feb;127(3):549-58
10631176
Stem Cells Dev. 2014 Dec 15;23 (24):3049-64
25144240
Methods Mol Med. 2006;121:125-48
16251740
PLoS One. 2015 Nov 11;10 (11):e0140902
26560325
Science. 1998 Dec 11;282(5396):2072-5
9851926
Development. 2010 Dec;137(24):4159-69
21098565
Development. 2005 May;132(9):2093-102
15788452
J Biol Chem. 1996 Jun 21;271(25):15292-7
8663044
Cell. 2005 Dec 2;123(5):917-29
16325584
J Endocrinol. 2000 May;165(2):443-56
10810308
Development. 2003 Oct;130(19):4527-37
12925581
Dev Biol. 2004 Jul 1;271(1):26-37
15196947
Endocrinology. 1992 Oct;131(4):1595-602
1396305
Mol Cell Biol. 2005 Dec;25(23):10479-91
16287860
Mol Reprod Dev. 2011 Jul;78(7):519-28
21710638
Eur Heart J. 1997 Jun;18(6):883-4
9183571
J Soc Gynecol Investig. 2005 Jan;12(1):2-13
15629664
Cell Cycle. 2012 Jul 1;11(13):2427-30
22659845
Dev Biol. 2007 Apr 15;304(2):567-78
17289015
Theriogenology. 2001 Jan 1;55(1):193-207
11198082