title: Regeneration of uterine horns in rats using collagen scaffolds loaded with human embryonic stem cell-derived endometrium-like cells.
abstract: A variety of diseases may lead to hysterectomies or uterine injuries, which may form a scar and lead to infertility. Due to the limitation of native materials, there are a few effective methods to treat such damages. Tissue engineering combines cell and molecular biology with materials and mechanical engineering to replace or repair damaged organs and tissues. The use of human embryonic stem cells (hESCs) as a donor cell source for the replacement therapy will require the development of simple and reliable cell differentiation protocols. This study aimed at efficiently generating endometrium-like cells from the hESCs and at using these cells with collagen scaffold to repair uterine damage. The hESCs were induced by co-culturing with endometrial stromal cells, and simultaneously added cytokines: epidermal growth factor (EGF), platelet-derived growth factor-b (PDGF-b), and E2. Expression of cell specific markers was analyzed by immunofluorescence and reverse trascription-polymerase chain reaction to monitor the progression toward an endometrium-like cell fate. After differentiation, the majority of cells (>80%) were positive for cytokeratin-7, and the expression of key transcription factors related to endometrial development, such as Wnt4, Wnt7a, Wnt5a, Hoxa11, and factors associated with endometrial epithelial cell function: Hoxa10, Intergrinβ3, LIF, ER, and PR were also detected. Then, we established the uterine full-thickness-injury rat models to test cell function in vivo. hESC-derived cells were dropped onto collagen scaffolds and transplanted into the animal model. Twelve weeks after transplantation, we discovered that the hESC-derived cells could survive and recover the structure and function of uterine horns in a rat model of severe uterine damage. The experimental system presented here provides a reliable protocol to produce endometrium-like cells from hESCs. Our results encourage the use of hESCs in cell-replacement therapy for severe uterine damage in future.
authors.affiliation: 1 Department of Obstetrics and Gynecology, Nanjing Drum Tower Hospital, Nanjing University Medical School , Nanjing, China .
authors.forename: Tianran
authors.initials: T
authors.lastname: Song
authors.forename: Xia
authors.initials: X
authors.lastname: Zhao
authors.forename: Haixiang
authors.initials: H
authors.lastname: Sun
authors.forename: Xin'an
authors.initials: X
authors.lastname: Li
authors.forename: Nacheng
authors.initials: N
authors.lastname: Lin
authors.forename: Lijun
authors.initials: L
authors.lastname: Ding
authors.forename: Jianwu
authors.initials: J
authors.lastname: Dai
authors.forename: Yali
authors.initials: Y
authors.lastname: Hu
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D004717:Endometrium / Q000166:cytology* / Q000637:transplantation; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D017207:Rats, Sprague-Dawley; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D015415:Biomarkers; D003094:Collagen
references.citation: Am J Pathol. 2003 Jul;163(1):295-301
references.pmid: 12819034
references.citation: Hum Reprod. 2007 May;22(5):1214-23
references.pmid: 17283036
references.citation: Biol Reprod. 2004 Jun;70(6):1738-50
references.pmid: 14766732
references.citation: Biol Reprod. 2010 Jun;82(6):1076-87
references.pmid: 20147733
references.citation: Biomaterials. 2011 Nov;32(32):8172-81
references.pmid: 21821282
references.citation: Hum Reprod. 2007 Nov;22(11):2903-11
references.pmid: 17872908
references.citation: Circ Res. 2013 Feb 1;112(3):523-33
references.pmid: 23371901
references.citation: Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2009 Mar-Apr;1(2):226-36
references.pmid: 20049793
references.citation: Rev Clin Esp. 2012 Sep;212(8):403-6
references.pmid: 22765959
references.citation: Fertil Steril. 2013 Nov;100(5):1358-63
references.pmid: 23830110
references.citation: Biol Reprod. 1998 Mar;58(3):627-32
references.pmid: 9510949
references.citation: Cell. 2004 Mar 19;116(6):769-78
references.pmid: 15035980
references.citation: Biochem Biophys Res Commun. 2009 Jan 30;379(1):16-21
references.pmid: 19084499
references.citation: J Minim Invasive Gynecol. 2007 Jan-Feb;14(1):15-22
references.pmid: 17218224
references.citation: Endocrinology. 1992 Apr;130(4):2373-85
references.pmid: 1312455
references.citation: Fertil Steril. 2014 Feb;101(2):587-94
references.pmid: 24355044
references.citation: J Exp Zool. 2001 Nov 1;290(6):616-23
references.pmid: 11748610
references.citation: Biol Reprod. 2009 Jun;80(6):1136-45
references.pmid: 19228591
references.citation: Semin Reprod Med. 2008 Jul;26(4):349-55
references.pmid: 18756412
references.citation: Annu Rev Cell Dev Biol. 2005;21:605-31
references.pmid: 16212509
references.citation: Biomaterials. 2004 May;25(11):2065-73
references.pmid: 14741621
references.citation: Curr Opin Organ Transplant. 2013 Apr;18(2):179-85
references.pmid: 23449347
references.citation: Obstet Gynecol Int. 2012;2012:853269
references.pmid: 22190959
references.citation: Stem Cells. 2007 Aug;25(8):2082-6
references.pmid: 17464086
references.citation: Tissue Eng Part B Rev. 2008 Jun;14(2):199-215
references.pmid: 18544016
references.citation: J Soc Gynecol Investig. 1997 May-Jun;4(3):160-6
references.pmid: 9258881
references.citation: Fertil Steril. 2002 Aug;78(2):221-33
references.pmid: 12137855
references.citation: Ann N Y Acad Sci. 2004 Dec;1034:152-65
references.pmid: 15731308
references.citation: J Clin Pathol. 2001 Jun;54(6):435-40
references.pmid: 11376015
references.citation: Int J Gynaecol Obstet. 2002 Mar;76(3):245-51
references.pmid: 11880127
references.citation: Dev Dyn. 2009 May;238(5):1118-30
references.pmid: 19384957
references.citation: Ann Afr Med. 2012 Apr-Jun;11(2):65-9
references.pmid: 22406663
references.citation: Respiration. 2013;85(1):3-10
references.pmid: 23257690
references.citation: Reprod Sci. 2007 Sep;14(6):524-33
references.pmid: 17959881
references.citation: Fertil Steril. 1994 Jan;61(1):1-17
references.pmid: 7507444
references.citation: Hum Reprod. 2008 Jan;23(1):139-43
references.pmid: 17981818
references.citation: Fertil Steril. 2013 Mar 1;99(3):605-6
references.pmid: 23714435
references.citation: Stem Cells. 2007 Nov;25(11):2820-6
references.pmid: 17656643
references.citation: Hum Reprod Update. 2007 Jan-Feb;13(1):87-101
references.pmid: 16960017
references.citation: Tissue Eng Part A. 2009 Oct;15(10):2927-35
references.pmid: 19290803
references.citation: Fertil Steril. 2013 Feb;99(2):470-6
references.pmid: 23084266
references.citation: Tissue Eng Part A. 2009 Jul;15(7):1611-8
references.pmid: 19061433
references.citation: Eur J Obstet Gynecol Reprod Biol. 2012 Feb;160(2):185-90
references.pmid: 22118886
references.citation: Zhonghua Nan Ke Xue. 2008 Dec;14(12):1083-9
references.pmid: 19157227