ABSTRACT
Recently, significant advances in our understanding of embryo implantation have been
made using animal models, notably in the mouse and nonhuman primates. However, the
determination of the molecular and cellular events that underpin the early stages
of implantation in the human remains an intractable problem, in part due to the inaccessibility
of early human implantation sites. In the absence of in vivo implantation sites, several
experimental in vitro model systems have been developed recently that mimic the different
stages of human embryo implantation that occur in vivo during the first few weeks
of pregnancy. These include solid-phase assays of blastocyst attachment and trophoblast
invasion, and two- and three-dimensional blastocyst-endometrial cell cocultures. An
important feature of such models is that they allow functional studies to be performed
and are not restricted to generating descriptive data. These models have the potential
to make an important contribution to the development of new therapeutic and diagnostic
strategies for implantation failure in the future, as well as toxicity testing. We
describe the strengths and weakness of the models and some of the advances that have
been made by the use of these in vitro models.
KEYWORDS
Implantation - in vitro - models - human - embryo
REFERENCES
1
Bulletti C, Flamigni C, Giacomucci E.
Reproductive failure due to spontaneous abortion and recurrent miscarriage.
Hum Reprod Update.
1996;
2(2)
118-136
2
HFEA .
Human Fertilisation and Embryo Authority. UK Clinic Success Rates.
Available at: http://www.hfea.gov.uk
Accessed June 5, 2007;
3
Kao L C, Tulac S, Lobo S et al..
Global gene profiling in human endometrium during the window of implantation.
Endocrinology.
2002;
143(6)
2119-2138
4
Achache H, Revel A.
Endometrial receptivity markers, the journey to successful embryo implantation.
Hum Reprod Update.
2006;
12(6)
731-746
5
Watson A J, Natale D R, Barcroft L C.
Molecular regulation of blastocyst formation.
Anim Reprod Sci.
2004;
82-83
583-592
6
Dominguez F, Pellicer A, Simon C.
Paracrine dialogue in implantation.
Mol Cell Endocrinol.
2002;
186(2)
175-181
7
Lessey B A, Damjanovich L, Coutifaris C, Castelbaum A, Albelda S M, Buck C A.
Integrin adhesion molecules in the human endometrium. Correlation with the normal
and abnormal menstrual cycle.
J Clin Invest.
1992;
90(1)
188-195
8
Kumar S, Zhu L J, Polihronis M et al..
Progesterone induces calcitonin gene expression in human endometrium within the putative
window of implantation.
J Clin Endocrinol Metab.
1998;
83(12)
4443-4450
9
Sherwin J R, Smith S K, Wilson A, Sharkey A M.
Soluble gp130 is up-regulated in the implantation window and shows altered secretion
in patients with primary unexplained infertility.
J Clin Endocrinol Metab.
2002;
87(8)
3953-3960
10
Fukuda M N, Sato T, Nakayama J et al..
Trophinin and tastin, a novel cell adhesion molecule complex with potential involvement
in embryo implantation.
Genes Dev.
1995;
9(10)
1199-1210
11
Campbell S, Swann H R, Seif M W, Kimber S J, Aplin J D.
Cell adhesion molecules on the oocyte and preimplantation human embryo.
Hum Reprod.
1995;
10(6)
1571-1578
12
Alikani M.
Epithelial cadherin distribution in abnormal human pre-implantation embryos.
Hum Reprod.
2005;
20(12)
3369-3375
13
Turpeenniemi-Hujanen T, Ronnberg L, Kauppila A, Puistola U.
Laminin in the human embryo implantation: analogy to the invasion by malignant cells.
Fertil Steril.
1992;
58(1)
105-113
14
Shimomura Y, Ando H, Furugori K et al..
Possible involvement of crosstalk cell-adhesion mechanism by endometrial CD26/dipeptidyl
peptidase IV and embryonal fibronectin in human blastocyst implantation.
Mol Hum Reprod.
2006;
12(8)
491-495
15
Dominguez F, Galan A, Martin J J, Remohi J, Pellicer A, Simon C.
Hormonal and embryonic regulation of chemokine receptors CXCR1, CXCR4, CCR5 and CCR2B
in the human endometrium and the human blastocyst.
Mol Hum Reprod.
2003;
9(4)
189-198
16
Chobotova K, Spyropoulou I, Carver J et al..
Heparin-binding epidermal growth factor and its receptor ErbB4 mediate implantation
of the human blastocyst.
Mech Dev.
2002;
119(2)
137-144
17
Wang H, Dey S K.
Roadmap to embryo implantation: clues from mouse models.
Nat Rev Genet.
2006;
7(3)
185-199
18
Stewart C L, Kaspar P, Brunet L J et al..
Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor.
Nature.
1992;
359(6390)
76-79
19
Ernst M, Inglese M, Waring P et al..
Defective gp130-mediated signal transducer and activator of transcription (STAT) signaling
results in degenerative joint disease, gastrointestinal ulceration, and failure of
uterine implantation.
J Exp Med.
2001;
194(2)
189-203
20
Bilinski P, Roopenian D, Gossler A.
Maternal IL-11Ralpha function is required for normal decidua and fetoplacental development
in mice.
Genes Dev.
1998;
12(14)
2234-2243
21
Lim H, Paria B C, Das S K et al..
Multiple female reproductive failures in cyclooxygenase 2-deficient mice.
Cell.
1997;
91(2)
197-208
22
Lee K Y, DeMayo F J.
Animal models of implantation.
Reproduction.
2004;
128(6)
679-695
23
Einspanier A, Lieder K, Einspanier R, Husen B.
The common marmoset monkey as a model for implantation and early pregnancy research.
Methods Mol Med.
2006;
121
111-121
24
Ghosh D, Sengupta J.
Target-oriented anti-implantation approaches for pregnancy interception: experiences
in the rhesus monkey model.
Contraception.
2005;
71(4)
294-301
25
Rosario G X, Modi D N, Sachdeva G, Manjramkar D D, Puri C P.
Morphological events in the primate endometrium in the presence of a preimplantation
embryo, detected by the serum preimplantation factor bioassay.
Hum Reprod.
2005;
20(1)
61-71
26
Fazleabas A T, Kim J J, Strakova Z.
Implantation: embryonic signals and the modulation of the uterine environment-a review.
Placenta.
2004;
25(suppl A)
S26-S31
27
Schultz J F, Mayernik L, Rout U K, Armant D R.
Integrin trafficking regulates adhesion to fibronectin during differentiation of mouse
peri-implantation blastocysts.
Dev Genet.
1997;
21(1)
31-43
28
Wang J, Armant D R.
Integrin-mediated adhesion and signaling during blastocyst implantation.
Cells Tissues Organs.
2002;
172(3)
190-201
29
Yelian F D, Yang Y, Hirata J D, Schultz J F, Armant D R.
Molecular interactions between fibronectin and integrins during mouse blastocyst outgrowth.
Mol Reprod Dev.
1995;
41(4)
435-448
30
Armant D R, Kaplan H A, Lennarz W J.
Fibronectin and laminin promote in vitro attachment and outgrowth of mouse blastocysts.
Dev Biol.
1986;
116(2)
519-523
31
Yelian F D, Edgeworth N A, Dong L J, Chung A E, Armant D R.
Recombinant entactin promotes mouse primary trophoblast cell adhesion and migration
through the Arg-Gly-Asp (RGD) recognition sequence.
J Cell Biol.
1993;
121(4)
923-929
32
Raab G, Kover K, Paria B C, Dey S K, Ezzell R M, Klagsbrun M.
Mouse preimplantation blastocysts adhere to cells expressing the transmembrane form
of heparin-binding EGF-like growth factor.
Development.
1996;
122(2)
637-645
33
Cavagna M, Mantese J C.
Biomarkers of endometrial receptivity-a review.
Placenta.
2003;
24(suppl B)
S39-S47
34
Lindenberg S, Hyttel P, Lenz S, Holmes P V.
Ultrastructure of the early human implantation in vitro.
Hum Reprod.
1986;
1(8)
533-538
35
Simon C, Mercader A, Garcia-Velasco J et al..
Coculture of human embryos with autologous human endometrial epithelial cells in patients
with implantation failure.
J Clin Endocrinol Metab.
1999;
84(8)
2638-2646
36
Simon C, Gimeno M J, Mercader A et al..
Embryonic regulation of integrins beta 3, alpha4, and alpha 1 in human endometrial
epithelial cells in vitro.
J Clin Endocrinol Metab.
1997;
82(8)
2607-2616
37
Gonzalez R R, Caballero-Campo P, Jasper M et al..
Leptin and leptin receptor are expressed in the human endometrium and endometrial
leptin secretion is regulated by the human blastocyst.
J Clin Endocrinol Metab.
2000;
85(12)
4883-4888
38
Meseguer M, Aplin J D, Caballero-Campo P et al..
Human endometrial mucin MUC1 is up-regulated by progesterone and down-regulated in
vitro by the human blastocyst.
Biol Reprod.
2001;
64(2)
590-601
39
Heneweer C, Kruse L H, Kindhauser F et al..
Adhesiveness of human uterine epithelial RL95-2 cells to trophoblast: rho protein
regulation.
Mol Hum Reprod.
2002;
8(11)
1014-1022
40
Tinel H, Denker H W, Thie M.
Calcium influx in human uterine epithelial RL95-2 cells triggers adhesiveness for
trophoblast-like cells. Model studies on signalling events during embryo implantation.
Mol Hum Reprod.
2000;
6(12)
1119-1130
41
White T E, Saltzman R A, Di Sant'Agnese P A, Keng P C, Sutherland R M, Miller R K.
Human choriocarcinoma (JAr) cells grown as multicellular spheroids.
Placenta.
1988;
9(6)
583-598
42
Heneweer C, Adelmann H G, Kruse L H, Denker H W, Thie M.
Human uterine epithelial RL95-2 cells reorganize their cytoplasmic architecture with
respect to Rho protein and F-actin in response to trophoblast binding.
Cells Tissues Organs.
2003;
175(1)
1-8
43
Thie M, Rospel R, Dettmann W et al..
Interactions between trophoblast and uterine epithelium: monitoring of adhesive forces.
Hum Reprod.
1998;
13(11)
3211-3219
44
Hess A P, Hamilton A E, Talbi S et al..
Decidual stromal cell response to paracrine signals from the trophoblast: amplification
of immune and angiogenic modulators.
Biol Reprod.
2007;
76(1)
102-117
45
Popovici R M, Betzler N K, Krause M S et al..
Gene expression profiling of human endometrial-trophoblast interaction in a coculture
model.
Endocrinology.
2006;
147(12)
5662-5675
46
Zeng B X, Fujiwara H, Sato Y et al..
Integrin alpha5 is involved in fibronectin-induced human extravillous trophoblast
invasion.
J Reprod Immunol.
2007;
73(1)
1-10
47
Spessotto P, Bulla R, Danussi C et al..
EMILIN1 represents a major stromal element determining human trophoblast invasion
of the uterine wall.
J Cell Sci.
2006;
119(pt 21)
4574-4584
48
Natanson-Yaron S, Anteby E Y, Greenfield C et al..
FGF 10 and Sprouty 2 modulate trophoblast invasion and branching morphogenesis.
Mol Hum Reprod.
2007;
13(7)
511-519
49
Hannan N J, Jones R L, White C A, Salamonsen L A.
The chemokines, CX3CL1, CCL14, and CCL4, promote human trophoblast migration at the
feto-maternal interface.
Biol Reprod.
2006;
74(5)
896-904
50
Briese J, Oberndorfer M, Patschenik C et al..
Osteopontin is colocalized with the adhesion molecule CEACAM1 in the extravillous
trophoblast of the human placenta and enhances invasion of CEACAM1-expressing placental
cells.
J Clin Endocrinol Metab.
2005;
90(9)
5407-5413
51
Pollheimer J, Loregger T, Sonderegger S et al..
Activation of the canonical wingless/T-cell factor signaling pathway promotes invasive
differentiation of human trophoblast.
Am J Pathol.
2006;
168(4)
1134-1147
52
Pongcharoen S, Somran J, Sritippayawan S et al..
Interleukin-17 expression in the human placenta.
Placenta.
2007;
28(1)
59-63
53
Bamberger A M, Minas V, Kalantaridou S N et al..
Corticotropin-releasing hormone modulates human trophoblast invasion through carcinoembryonic
antigen-related cell adhesion molecule-1 regulation.
Am J Pathol.
2006;
168(1)
141-150
54
Huber A V, Saleh L, Bauer S, Husslein P, Knofler M.
TNFalpha-mediated induction of PAI-1 restricts invasion of HTR-8/SVneo trophoblast
cells.
Placenta.
2006;
27(2-3)
127-136
55
Tartakover-Matalon S, Cherepnin N, Kuchuk M et al..
Impaired migration of trophoblast cells caused by simvastatin is associated with decreased
membrane IGF-I receptor, MMP2 activity and HSP27 expression.
Hum Reprod.
2007;
22(4)
1161-1167
56
Mandl M, Ghaffari-Tabrizi N, Haas J, Nohammer G, Desoye G.
Differential glucocorticoid effects on proliferation and invasion of human trophoblast
cell lines.
Reproduction.
2006;
132(1)
159-167
57
Carver J, Martin K, Spyropoulou I, Barlow D, Sargent I, Mardon H.
An in-vitro model for stromal invasion during implantation of the human blastocyst.
Hum Reprod.
2003;
18(2)
283-290
58
Martin K L, Barlow D H, Sargent I L.
Heparin-binding epidermal growth factor significantly improves human blastocyst development
and hatching in serum-free medium.
Hum Reprod.
1998;
13(6)
1645-1652
59
Jones M C, Fusi L, Higham J H et al..
Regulation of the SUMO pathway sensitizes differentiating human endometrial stromal
cells to progesterone.
Proc Natl Acad Sci USA.
2006;
103(44)
16272-16277
60
Cervero A, Dominguez F, Horcajadas J A, Quinonero A, Pellicer A, Simon C.
Embryonic adhesion is not affected by endometrial leptin receptor gene silencing.
Fertil Steril.
2007;
, March 2 (Epub ahead of print)
61
Shiokawa S, Sakai K, Akimoto Y et al..
Function of the small guanosine triphosphate-binding protein RhoA in the process of
implantation.
J Clin Endocrinol Metab.
2000;
85(12)
4742-4749
62
Madan P, Rose K, Watson A J.
Na/K-ATPase beta1 subunit expression is required for blastocyst formation and normal
assembly of trophectoderm tight junction-associated proteins.
J Biol Chem.
2007;
282(16)
12127-12134
63
Georgiades P, Cox B, Gertsenstein M, Chawengsaksophak K, Rossant J.
Trophoblast-specific gene manipulation using lentivirus-based vectors.
Biotechniques.
2007;
42(3)
317-318
64
Bigsby R M.
Control of growth and differentiation of the endometrium: the role of tissue interactions.
Ann N Y Acad Sci.
2002;
955
110-118
65
Punyadeera C, Verbost P, Groothuis P.
Oestrogen and progestin responses in human endometrium.
J Steroid Biochem Mol Biol.
2003;
84(4)
393-410
66
Bentin-Ley U, Pedersen B, Lindenberg S, Larsen J F, Hamberger L, Horn T.
Isolation and culture of human endometrial cells in a three-dimensional culture system.
J Reprod Fertil.
1994;
101(2)
327-332
67
Kim M R, Park D W, Lee J H et al..
Progesterone-dependent release of transforming growth factor-beta1 from epithelial
cells enhances the endometrial decidualization by turning on the Smad signalling in
stromal cells.
Mol Hum Reprod.
2005;
11(11)
801-808
68
Igarashi T M, Bruner-Tran K L, Yeaman G R et al..
Reduced expression of progesterone receptor-B in the endometrium of women with endometriosis
and in cocultures of endometrial cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.
Fertil Steril.
2005;
84(1)
67-74
69
Blauer M, Heinonen P K, Martikainen P M, Tomas E, Ylikomi T.
A novel organotypic culture model for normal human endometrium: regulation of epithelial
cell proliferation by estradiol and medroxyprogesterone acetate.
Hum Reprod.
2005;
20(4)
864-871
70
Pierro E, Minici F, Alesiani O et al..
Stromal-epithelial interactions modulate estrogen responsiveness in normal human endometrium.
Biol Reprod.
2001;
64(3)
831-838
71
Classen-Linke I, Kusche M, Knauthe R, Beier H M.
Establishment of a human endometrial cell culture system and characterization of its
polarized hormone responsive epithelial cells.
Cell Tissue Res.
1997;
287(1)
171-185
72
Arnold J T, Kaufman D G, Seppala M, Lessey B A.
Endometrial stromal cells regulate epithelial cell growth in vitro: a new co-culture
model.
Hum Reprod.
2001;
16(5)
836-845
73
Park D W, Choi D S, Ryu H S, Kwon H C, Joo H, Min C K.
A well-defined in vitro three-dimensional culture of human endometrium and its applicability
to endometrial cancer invasion.
Cancer Lett.
2003;
195(2)
185-192
74
Bentin-Ley U, Horn T, Sjogren A, Sorensen S, Falck Larsen J, Hamberger L.
Ultrastructure of human blastocyst-endometrial interactions in vitro.
J Reprod Fertil.
2000;
120(2)
337-350
75
Petersen A, Bentin-Ley U, Ravn V et al..
The antiprogesterone Org 31710 inhibits human blastocyst-endometrial interactions
in vitro.
Fertil Steril.
2005;
83(suppl 1)
1255-1263
Helen Mardon
Nuffield Department of Obstetrics and Gynaecology, University of Oxford
Level 3, The Women's Centre, John Radcliffe Hospital, Headington, Oxford, OX3 9DU
United Kingdom
eMail: helen.mardon@obs-gyn.ox.ac.uk