107
In this study, we investigated the supportive role of stromal cells in embryonic stem cell (ESC) culture. We sought to develop a method for using BMSCs as feeder cells for the establishment and culture of stem cells, based on the ease of manipulation of BMSCs as feeder cells and their expression of the LIF gene.
As described in Chapter 3, we determined that several different BMSC lines secreted between 42 and 75 pg/ml of LIF.
LIF plays a pivotal role as an extrinsic factor for ES self-renewal.
LIF binds to heterodimer receptors, the LIF receptor (LIFR) and gp130. After binding, tyrosine kinase Janus kinase (JAK) phosphorylates tyrosine residues of LIFR and GP130. STAT3 is then activated, and translocates into the nucleus where it functions as a transcription factor for several target genes inclduing myc, fos and klf4 (Niwa et al., 1998). The secretion of LIF by BMSC lines may explain why ESC maintenance does not depend on exogenous LIF. The use of BMSCs sub-passaged more than 20 times minimizes LIF enrichment and contributes to improving the efficiency of the stem cell co-culture system. No significant difference in morphology or the expression of several stemness-related genes was detected between BMSC and MEF co-cultures. These results demonstrate that BMSCs can be a viable substitute for MEF as feeder cells for mouse ESCs, which may further optimize the stem cell culture systems while reducing the need for animal euthanasia. In previous studies (Kawasaki et al., 2000; Shintani et al., 2008), ESCs were found to efficiently differentiate into dopaminergic neurons when cultured on a BMSC feeder layer in neuronal differentiation
108
medium. Therefore, further studies are required to determine aspects of the differentiation properties of ESCs cultured with BMSC and MEF feeders.
In Chapters 4 and 5, we investigated the properties of stromal cell differentiation. We examined the role of the ERK1/2 signaling pathways during the differentiation of uterine stromal cells. ERK1/2 is a member of the MAPK pathway and is implicated as a critical regulator of cellular proliferation and differentiation in multiple organ systems. ERK1/2 is activated by the upstream Ras/Raf/MEK signaling cascade via phosphorylation on residues threonine 202 and tyrosine 204.
This activated form of ERK1/2 (phospho-ERK1/2) can subsequently phosphorylate various transcription factors to regulate cellular proliferation, differentiation, apoptosis and inflammation (Roskoski, 2012).
In the results of Chapter 4, phospho-ERK1/2 is first seen at 4.5 dpc in the decidualized cells surrounding the embryo in mice.
Its expression then expands to include the primary and secondary decidual zones at 5.5 and 7.5 dpc, respectively. Our real-time PCR results show that the known ERK1/2 target genes, Fos, Stat1, Ubtf, Elk1, Serpine1 and Msk1 are also significantly up-regulated on Day 5.5 at the implantation sites.
In hESCs, we found that ERK1/2 is highly expressed during decidualization. Inhibition of phospho-ERK1/2 suppresses decidualization of hESCs, as evidenced by decreases in
109
expression of the decidual marker genes (IGFBP1 and PRL). Our evidence also suggests that activation of ERK1/2 during decidualization results in the phosphorylation of C/EBPβ, an important gene previously shown to be implicated in the regulation of decidualization. These findings have direct implications for our understanding of implantation and may reveal new therapeutic targets for the treatment of endometrial pathologies and infertility.
Future studies are needed to determine the precise mechanisms by which ERK1/2 regulates its target gene expression for localized and systemic responses through in vivo ChIP-seq analyses that will identify ERK1/2 binding sites in the mouse uterus in vivo. Through these future studies, new knowledge will be gained to aid not only the development of novel treatments of female infertility, but also for the treatment of uterine disorders, such as endometriosis and endometrial cancer, which are associated with altered ERK1/2 signaling.
In conclusion, we have verified that stromal cells can be utilized as feeder cells for the successful culture of mouse embryonic stem cells, and have investigated signaling pathways activated during stromal cell differentiation. The results obtained from this study have implications for the understanding of stromal cells and their relevance in a number of human diseases.
110
References
A EM. 2007. Isolation and propagation of mouse embryonic fibroblasts and preparation of mouse embryonic feeder layer cells. Curr Protoc Stem Cell Biol Chapter 1:Unit1C 3.
Aghajanova L, Giudice LC. 2011. Molecular evidence for differences in endometrium in severe versus mild endometriosis. Reprod Sci 18:229-251.
Alborzi S, Momtahan M, Parsanezhad ME, Dehbashi S, Zolghadri J, Alborzi S. 2004. A prospective, randomized study comparing laparoscopic ovarian cystectomy versus fenestration and coagulation in patients with endometriomas. Fertil Steril 82:1633-1637.
Ansell JD, Barlow PW, McLaren A. 1974. Binucleate and polyploid cells in the decidua of the mouse. J Embryol Exp Morphol 31:223-227.
Armstrong L, Lako M, Lincoln J, Cairns PM, Hole N. 2000. mTert expression correlates with telomerase activity during the differentiation of murine embryonic stem cells. Mech Dev 97:109-116.
Arnold JT, Kaufman DG, Seppala M, Lessey BA. 2001.
Endometrial stromal cells regulate epithelial cell growth in vitro: a new co-culture model. Hum Reprod 16:836-845.
Aubert J, Dunstan H, Chambers I, Smith A. 2002. Functional gene screening in embryonic stem cells implicates Wnt antagonism in neural differentiation. Nat Biotechnol
111
Avruch J, Khokhlatchev A, Kyriakis JM, Luo Z, Tzivion G, Vavvas D, Zhang XF. 2001. Ras activation of the Raf kinase:
tyrosine kinase recruitment of the MAP kinase cascade.
Recent Prog Horm Res 56:127-155.
Bagchi IC, Li Q, Cheon YP, Mantena SR, Kannan A, Bagchi MK.
2005. Use of the progesterone receptor antagonist RU 486 to identify novel progesterone receptor-regulated pathways in implantation. Semin Reprod Med 23:38-45.
Bain G, Kitchens D, Yao M, Huettner JE, Gottlieb DI. 1995.
Embryonic stem cells express neuronal properties in vitro.
Dev Biol 168:342-357.
Baker DJ, Nagy F, Nieder GL. 1992. Localization of c-fos-like proteins in the mouse endometrium during the peri-implantation period. Biol Reprod 47:492-501.
Baksh D, Song L, Tuan RS. 2004. Adult mesenchymal stem cells:
characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med 8:301-316.
Banerjee P, Sapru K, Strakova Z, Fazleabas AT. 2009. Chorionic gonadotropin regulates prostaglandin E synthase via a phosphatidylinositol 3-kinase-extracellular regulatory kinase pathway in a human endometrial epithelial cell line:
implications for endometrial responses for embryo implantation. Endocrinology 150:4326-4337.
112
Blumenkrantz MJ, Gallagher N, Bashore RA, Tenckhoff H. 1981.
Retrograde menstruation in women undergoing chronic peritoneal dialysis. Obstet Gynecol 57:667-670.
Boeuf H, Hauss C, Graeve FD, Baran N, Kedinger C. 1997.
Leukemia inhibitory factor-dependent transcriptional activation in embryonic stem cells. J Cell Biol 138:1207-1217.
Boiani M, Scholer HR. 2005. Regulatory networks in embryo-derived pluripotent stem cells. Nat Rev Mol Cell Biol 6:872-884.
Boonyaratanakornkit V, Bi Y, Rudd M, Edwards DP. 2008. The role and mechanism of progesterone receptor activation of extra-nuclear signaling pathways in regulating gene transcription and cell cycle progression. Steroids 73:922-928.
Bradley A, Evans M, Kaufman MH, Robertson E. 1984. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature 309:255-256.
Brook FA, Gardner RL. 1997. The origin and efficient derivation of embryonic stem cells in the mouse. Proc Natl Acad Sci U S A 94:5709-5712.
Brosens JJ, Hayashi N, White JO. 1999. Progesterone receptor regulates decidual prolactin expression in differentiating human endometrial stromal cells. Endocrinology 140:4809-4820.
Brosens JJ, Parker MG, McIndoe A, Pijnenborg R, Brosens IA.
2009. A role for menstruation in preconditioning the uterus for successful pregnancy. Am J Obstet Gynecol
113
200:615 e611-616.Bryja V, Bonilla S, Cajanek L, Parish CL, Schwartz CM, Luo Y, Rao MS, Arenas E. 2006. An efficient method for the derivation of mouse embryonic stem cells. Stem Cells 24:844-849.
Buchwalter G, Gross C, Wasylyk B. 2004. Ets ternary complex transcription factors. Gene 324:1-14.
Buck M, Poli V, van der Geer P, Chojkier M, Hunter T. 1999.
Phosphorylation of rat serine 105 or mouse threonine 217 in C/EBP beta is required for hepatocyte proliferation induced by TGF alpha. Molecular cell 4:1087-1092.
Buehr M, Smith A. 2003. Genesis of embryonic stem cells. Philos Trans R Soc Lond B Biol Sci 358:1397-1402; discussion 1402.
Bulun SE, Zeitoun KM, Takayama K, Simpson E, Sasano H. 2000.
Aromatase as a therapeutic target in endometriosis.
Trends in Endocrinology and Metabolism 11:22-27.
Burney RO, Talbi S, Hamilton AE, Vo KC, Nyegaard M, Nezhat CR, Lessey BA, Giudice LC. 2007. Gene expression analysis of endometrium reveals progesterone resistance and candidate susceptibility genes in women with endometriosis. Endocrinology 148:3814-3826.
Cantley LC. 2002. The phosphoinositide 3-kinase pathway.
Science 296:1655-1657.
Chalmers CJ, Gilley R, March HN, Balmanno K, Cook SJ. 2007.
The duration of ERK1/2 activity determines the activation of c-Fos and Fra-1 and the composition and quantitative transcriptional output of AP-1. Cell Signal 19:695-704.
114
Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, Smith A. 2003. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells.
Cell 113:643-655.
Chambers I, Smith A. 2004. Self-renewal of teratocarcinoma and embryonic stem cells. Oncogene 23:7150-7160.
Chen RH, Sarnecki C, Blenis J. 1992. Nuclear localization and regulation of erk- and rsk-encoded protein kinases. Mol Cell Biol 12:915-927.
Cheng J, Dutra A, Takesono A, Garrett-Beal L, Schwartzberg PL.
2004. Improved generation of C57BL/6J mouse embryonic stem cells in a defined serum-free media. Genesis 39:100-104.
Christian M, Pohnke Y, Kempf R, Gellersen B, Brosens JJ. 2002.
Functional association of PR and CCAAT/enhancer-binding protein beta isoforms: promoter-dependent cooperation between PR-B and liver-enriched inhibitory protein, or liver-enriched activatory protein and PR-A in human endometrial stromal cells. Mol Endocrinol 16:141-154.
Conneely OM, Maxwell BL, Toft DO, Schrader WT, O'Malley BW.
1987. The A and B forms of the chicken progesterone receptor arise by alternate initiation of translation of a unique mRNA. Biochem Biophys Res Commun 149:493-501.
Conneely OM, Mulac-Jericevic B, Lydon JP. 2003.
Progesterone-dependent regulation of female reproductive activity by two distinct progesterone
115
receptor isoforms. Steroids 68:771-778.
Conner DA. 2001. Mouse embryo fibroblast (MEF) feeder cell preparation. Curr Protoc Mol Biol Chapter 23:Unit 23 22.
Crews CM, Alessandrini A, Erikson RL. 1992. The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product. Science 258:478-480.
Cui G, Qi Z, Guo X, Qin J, Gui Y, Cai Z. 2009. Rat bone marrow derived mesenchymal progenitor cells support mouse ES cell growth and germ-like cell differentiation. Cell Biol Int 33:434-441.
Cunha GR, Bigsby RM, Cooke PS, Sugimura Y. 1985. Stromal-epithelial interactions in adult organs. Cell Differ 17:137-148.
Daly DC, Maslar IA, Riddick DH. 1983. Prolactin production during in vitro decidualization of proliferative endometrium. Am J Obstet Gynecol 145:672-678.
de Ziegler D, Fanchin R, de Moustier B, Bulletti C. 1998. The hormonal control of endometrial receptivity: estrogen (E2) and progesterone. J Reprod Immunol 39:149-166.
Dey SK, Lim H, Das SK, Reese J, Paria BC, Daikoku T, Wang H.
2004. Molecular cues to implantation. Endocr Rev 25:341-373.
Dimitriadis E, Stoikos C, Tan YL, Salamonsen LA. 2006.
Interleukin 11 signaling components signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) regulate human endometrial stromal cell differentiation. Endocrinology 147:3809-3817.
116
Dimitriadis E, White CA, Jones RL, Salamonsen LA. 2005.
Cytokines, chemokines and growth factors in endometrium related to implantation. Hum Reprod Update 11:613-630.
Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R. 1985.
The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol 87:27-45.
Downing GJ, Battey JF, Jr. 2004. Technical assessment of the first 20 years of research using mouse embryonic stem cell lines. Stem Cells 22:1168-1180.
Duncan SA, Zhong Z, Wen ZL, Darnell JE. 1997. STAT signaling is active during early mammalian development. Dev Dynam 208:190-198.
Dyer MA, Farrington SM, Mohn D, Munday JR, Baron MH. 2001.
Indian hedgehog activates hematopoiesis and vasculogenesis and can respecify prospective neurectodermal cell fate in the mouse embryo.
Development 128:1717-1730.
Eskenazi B, Warner ML. 1997. Epidemiology of endometriosis.
Obstet Gynecol Clin North Am 24:235-258.
Evans MJ, Kaufman MH. 1981. Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154-156.
Fairchild PJ, Brook FA, Gardner RL, Graca L, Strong V, Tone Y, Tone M, Nolan KF, Waldmann H. 2000. Directed differentiation of dendritic cells from mouse embryonic
117
stem cells. Curr Biol 10:1515-1518.
Fan HY, Liu Z, Johnson PF, Richards JS. 2011.
CCAAT/enhancerbinding proteins (C/EBP)alpha and -beta are essential for ovulation, luteinization, and the expression of key target genes. Mol Endocrinol 25:253-268.
Fan HY, Liu Z, Shimada M, Sterneck E, Johnson PF, Hedrick SM, Richards JS. 2009. MAPK3/1 (ERK1/2) in ovarian granulosa cells are essential for female fertility. Science 324:938-941.
Fazleabas AT, Strakova Z. 2002. Endometrial function: cell specific changes in the uterine environment. Mol Cell Endocrinol 186:143-147.
Firulli AB. 2003. A HANDful of questions: the molecular biology of the heart and neural crest derivatives (HAND)-subclass of basic helix-loop-helix transcription factors.
Gene 312:27-40.
Franco HL, Dai D, Lee KY, Rubel CA, Roop D, Boerboom D, Jeong JW, Lydon JP, Bagchi IC, Bagchi MK, DeMayo FJ. 2011.
WNT4 is a key regulator of normal postnatal uterine development and progesterone signaling during embryo implantation and decidualization in the mouse. FASEB J 25:1176-1187.
Franco HL, Jeong JW, Tsai SY, Lydon JP, DeMayo FJ. 2008. In vivo analysis of progesterone receptor action in the uterus during embryo implantation. Semin Cell Dev Biol 19:178-186.
Gao J, Mazella J, Seppala M, Tseng L. 2001. Ligand activated hPR
118
modulates the glycodelin promoter activity through the Sp1 sites in human endometrial adenocarcinoma cells. Mol Cell Endocrinol 176:97-102.
Gellersen B, Brosens IA, Brosens JJ. 2007. Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med 25:445-453.
Ginis I, Luo Y, Miura T, Thies S, Brandenberger R, Gerecht-Nir S, Amit M, Hoke A, Carpenter MK, Itskovitz-Eldor J, Rao MS. 2004. Differences between human and mouse embryonic stem cells. Dev Biol 269:360-380.
Giudice LC. 2010. Clinical practice. Endometriosis. N Engl J Med 362:2389-2398.
Giudice LC, Telles TL, Lobo S, Kao L. 2002. The molecular basis for implantation failure in endometriosis: on the road to discovery. Ann N Y Acad Sci 955:252-264; discussion 293-255, 396-406.
Gizard F, Robillard R, Gervois P, Faucompre A, Revillion F, Peyrat JP, Hum WD, Staels B. 2005. Progesterone inhibits human breast cancer cell growth through transcriptional upregulation of the cyclin-dependent kinase inhibitor p27Kip1 gene. FEBS Lett 579:5535-5541.
Goldstein DP, deCholnoky C, Emans SJ, Leventhal JM. 1980.
Laparoscopy in the diagnosis and management of pelvic pain in adolescents. J Reprod Med 24:251-256.
Hahnel AC, Rappolee DA, Millan JL, Manes T, Ziomek CA, Theodosiou NG, Werb Z, Pedersen RA, Schultz GA. 1990.
Two alkaline phosphatase genes are expressed during early development in the mouse embryo. Development
119
110:555-564.Hastings JM, Jackson KS, Mavrogianis PA, Fazleabas AT. 2006.
The Estrogen Early Response Gene FOS Is Altered in a Baboon Model of Endometriosis. Biol Reprod 75:176-182.
Heinrich PC, Behrmann I, Haan S, Hermanns HM, Muller-Newen G, Schaper F. 2003. Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochem J 374:1-20.
Hsu YL, Hou MF, Kuo PL, Huang YF, Tsai EM. 2012. Breast tumor-associated osteoblast-derived CXCL5 increases cancer progression by ERK/MSK1/Elk-1/Snail signaling pathway. Oncogene.
Irwin JC, Kirk D, King RJ, Quigley MM, Gwatkin RB. 1989.
Hormonal regulation of human endometrial stromal cells in culture: an in vitro model for decidualization. Fertil Steril 52:761-768.
Irwin JC, Utian WH, Eckert RL. 1991. Sex steroids and growth factors differentially regulate the growth and differentiation of cultured human endometrial stromal cells.
Endocrinology 129:2385-2392.
Ismail PM, Li J, DeMayo FJ, O'Malley BW, Lydon JP. 2002. A novel LacZ reporter mouse reveals complex regulation of the progesterone receptor promoter during mammary gland development. Mol Endocrinol 16:2475-2489.
Jeong JW, Kwak I, Lee KY, Kim TH, Large MJ, Stewart CL, Kaestner KH, Lydon JP, DeMayo FJ. 2010. Foxa2 Is Essential for Mouse Endometrial Gland Development and Fertility. Biology of Reproduction 83:396-403.
120
Jeong JW, Lee KY, Kwak I, White LD, Hilsenbeck SG, Lydon JP, DeMayo FJ. 2005. Identification of murine uterine genes regulated in a ligand-dependent manner by the progesterone receptor. Endocrinology 146:3490-3505.
Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM. 2002. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418:41-49.
Jirmanova L, Afanassieff M, Gobert-Gosse S, Markossian S, Savatier P. 2002. Differential contributions of ERK and PI3-kinase to the regulation of cyclin D1 expression and to the control of the G1/S transition in mouse embryonic stem cells. Oncogene 21:5515-5528.
Kania G, Corbeil D, Fuchs J, Tarasov KV, Blyszczuk P, Huttner WB, Boheler KR, Wobus AM. 2005. Somatic stem cell marker prominin-1/CD133 is expressed in embryonic stem cell-derived progenitors. Stem Cells 23:791-804.
Kannan A, Fazleabas AT, Bagchi IC, Bagchi MK. 2010. The profiling of endometrium from women with endometriosis reveals candidate genes for disease-based implantation failure and infertility. Endocrinology 144:2870-2881.
Karatza C, Shall S. 1984. The reproductive potential of normal
121
mouse embryo fibroblasts during culture in vitro. J Cell Sci 66:401-409.
Karpovich N, Klemmt P, Hwang JH, McVeigh JE, Heath JK, Barlow DH, Mardon HJ. 2005. The production of interleukin-11 and decidualization are compromised in endometrial stromal cells derived from patients with infertility. J Clin Endocrinol Metab 90:1607-1612.
Kastner P, Krust A, Turcotte B, Stropp U, Tora L, Gronemeyer H, Chambon P. 1990. Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. Embo J 9:1603-1614.
Kawasaki H, Mizuseki K, Nishikawa S, Kaneko S, Kuwana Y, Nakanishi S, Nishikawa SI, Sasai Y. 2000. Induction of midbrain dopaminergic neurons from ES cells by stromal cell-derived inducing activity. Neuron 28:31-40.
Kawase E, Suemori H, Takahashi N, Okazaki K, Hashimoto K, Nakatsuji N. 1994. Strain difference in establishment of mouse embryonic stem (ES) cell lines. Int J Dev Biol 38:385-390.
Keller GM. 1995. In vitro differentiation of embryonic stem cells.
Curr Opin Cell Biol 7:862-869.
Kern H, Zevnik B. 2009. ES cell line establishment. Methods Mol Biol 530:187-204.
Kielman MF, Rindapaa M, Gaspar C, van Poppel N, Breukel C, van Leeuwen S, Taketo MM, Roberts S, Smits R, Fodde R.
2002. Apc modulates embryonic stem-cell differentiation by controlling the dosage of beta-catenin signaling. Nat
122
Genet 32:594-605.Kim IW, Gong SP, Yoo CR, Choi JH, Kim DY, Lim JM. 2009.
Derivation of developmentally competent oocytes by the culture of preantral follicles retrieved from adult ovaries:
maturation, blastocyst formation, and embryonic stem cell transformation. Fertil Steril 92:1716-1724.
Kim J, Denu RA, Dollar BA, Escalante LE, Kuether JP, Callander NS, Asimakopoulos F, Hematti P. 2012. Macrophages and mesenchymal stromal cells support survival and proliferation of multiple myeloma cells. Br J Haematol 158:336-346.
Kim JJ, Jaffe RC, Fazleabas AT. 1998. Comparative studies on the in vitro decidualization process in the baboon (Papio anubis) and human. Biol Reprod 59:160-168.
Klemmt PA, Carver JG, Kennedy SH, Koninckx PR, Mardon HJ.
2006. Stromal cells from endometriotic lesions and endometrium from women with endometriosis have reduced decidualization capacity. Fertil Steril 85:564-572.
Kosako H, Gotoh Y, Matsuda S, Ishikawa M, Nishida E. 1992.
Xenopus MAP kinase activator is a serine/threonine/tyrosine kinase activated by threonine phosphorylation. Embo J 11:2903-2908.
Kramer J, Hegert C, Guan K, Wobus AM, Muller PK, Rohwedel J.
2000. Embryonic stem cell-derived chondrogenic differentiation in vitro: activation by BMP-2 and BMP-4.
Mech Dev 92:193-205.
Kraus WL, Montano MM, Katzenellenbogen BS. 1993. Cloning of the rat progesterone receptor gene 5'-region and
123
identification of two functionally distinct promoters. Mol Endocrinol 7:1603-1616.
Kristensen DM, Kalisz M, Nielsen JH. 2005. Cytokine signalling in embryonic stem cells. APMIS 113:756-772.
Kurihara I, Lee DK, Petit FG, Jeong J, Lee K, Lydon JP, DeMayo FJ, Tsai MJ, Tsai SY. 2007. COUP-TFII mediates progesterone regulation of uterine implantation by controlling ER activity. PLoS Genet 3:e102.
Kyriakis JM, App H, Zhang XF, Banerjee P, Brautigan DL, Rapp UR, Avruch J. 1992. Raf-1 activates MAP kinase-kinase.
Nature 358:417-421.
Laird SM, Tuckerman EM, Li TC. 2006. Cytokine expression in the endometrium of women with implantation failure and recurrent miscarriage. Reprod Biomed Online 13:13-23.
Leahy A, Xiong JW, Kuhnert F, Stuhlmann H. 1999. Use of developmental marker genes to define temporal and spatial patterns of differentiation during embryoid body formation. J Exp Zool 284:67-81.
Lee JH, Kim TH, Oh SJ, Yoo JY, Akira S, Ku BJ, Lydon JP, Jeong JW. 2013. Signal transducer and activator of transcription-3 (Stat3) plays a critical role in implantation via progesterone receptor in uterus. FASEB J.
Lee JH, Lee EJ, Lee CH, Park JH, Han JY, Lim JM. 2009.
Requirement of leukemia inhibitory factor for establishing and maintaining embryonic stem cells in mice. Fertil Steril 92:1133-1140.
Lee JT, Jr., McCubrey JA. 2002. The Raf/MEK/ERK signal transduction cascade as a target for chemotherapeutic
124
intervention in leukemia. Leukemia 16:486-507.
Lee K, Jeong J, Kwak I, Yu CT, Lanske B, Soegiarto DW, Toftgard R, Tsai MJ, Tsai S, Lydon JP, DeMayo FJ. 2006.
Indian hedgehog is a major mediator of progesterone signaling in the mouse uterus. Nat Genet 38:1204-1209.
Lee KY, DeMayo FJ. 2004. Animal models of implantation.
Reproduction 128:679-695.
Lee KY, Jeong JW, Wang JR, Ma LJ, Martin JF, Tsai SY, Lydon JP, DeMayo FJ. 2007. Bmp2 is critical for the murine uterine decidual response. Molecular and Cellular Biology 27:5468-5478.
Lee ST, Yun JI, Jo YS, Mochizuki M, van der Vlies AJ, Kontos S, Ihm JE, Lim JM, Hubbell JA. 2010. Engineering integrin
Lee ST, Yun JI, Jo YS, Mochizuki M, van der Vlies AJ, Kontos S, Ihm JE, Lim JM, Hubbell JA. 2010. Engineering integrin