Estrogen treatment enhances neurogenic differentiation of human adipose derived stem cells in vitro

Document Type : Original Article


1 Department of Anatomical Sciences and Molecular Biology, Isfahan University of Medical Sciences, Isfahan, Iran

2 Molecular Parasitology Laboratory, Pasteur Institute of Iran, Tehran, Iran

3 Department of Biology, Islamic Azad University, Science and Research Branch, Tehran, Iran

4 Department of Genetic, Isfahan University of Medical Sciences, Isfahan, Iran


Objective(s):Estrogen is a sexual hormone that has prominent effects on reproductive and non-reproductive tissues. The aim of this study is to evaluate the effects of estrogen on the proliferation and neural differentiation of human adipose derived stem cells (ADSCs) during neurogenic differentiation.
Materials and Methods: Isolated human ADSCs were trans-differentiated in neural induction medium containing neurobasal medium, N2 and B27 with or without 17β-estradiol (E2) treatment. Proliferation rate and neural differentiation of human ADSCs were assessed using MTT assay, immunostaining and real time RT- PCR analysis, respectively. 
Results: Analysis of data show that estradiol treatment can significantly increase proliferation rate of differentiated cells (P<0.05). Immunocytochemical and real time RT-PCR analysis revealed that the expression of precursor and mature neuronal markers (nestin and MAP2) was significantly higher in the E2 treated cell cultures when compared to the untreated cell cultures (P<0.05).
Conclusion: According to our findings, estrogen can promote proliferation and neuronal differentiation of human ADSCs.


1. Kang SK, Lee DH, Bae YC, Kim HK, Baik SY, Jung JS. Improvement of neurological de3 cits by intracerebral transplant of human adipose-derived stem cells after ischemia in rats. Exp Neurol 2003; 83: 355–366.
2. Kang SK, Shin MJ, Jung JS, Kim YG, Kim CH. Autologous adipose tissue-derived stromal cells for treatment of spinal cord injury. Stem Cells Dev 2006; 15: 583–594.
3. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001; 7: 211-228.
4. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002; 13: 4279-4295.
5. Kern S, Eichler H, Stoeve J, Klüter H, Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood or adipose tissue. Stem Cells 2006; 24:1294–1301.
6. Brann DW, Dhandapani K, Wakade C, Mahesh VB, Khan MM. Neurotrophic and neuroprotective actions of estrogen: basic mechanisms and clinical implications. Steroids 2007; 72: 381–405.
7. Garcia-Segura LM, Azcoitia I, DonCarlos LL. DonCarlos, Neuroprotection by estradiol. Prog. Neurobiol. 2001; 63: 29-60.
8. Garcia-Segura LM, Veiga S, Sierra A, Melcangi RC, Azcoitia I. Aromatase: a neuroprotective enzyme. Prog. Neurobiol. 2003; 71: 31–41.
9. Suzuki S, Brown CM, Wise PM.Mechanisms of neuroprotection by estrogen. Endocrine 2006; 29: 209–215.
10. Suzuki S, Gerhold LM, Böttner M, Rau SW, Dela Cruz C, Yang E, Zhu H, Yu J, Cashion AB, Kindy MS, Merchenthaler I, Gage FH, Wise PM.Estradiol enhances neurogenesis following ischemic stroke through estrogen receptors alpha and beta, J. Comp. Neurol. 2007; 500: 1064–1075.
11. Brannvall K, Korhonen L, Lindholm D. Estrogen-receptor-dependent regulation of neural stem cell proliferation and differentiation. Mol Cell Neurosci 2002; 21:512–520.
12. Singh M, Meyer EM, MillardWJ, Simpkins JW. Ovariectomy reduces ChAT activity and NGF mRNA levels in the frontal cortex and hippocampus of the female Sprague–Dawley rat. Soc Neurosci Abstr 1993; 19:254.
13. Singh M, Meyer EM, Simpkins JW. The effect of ovariectomy and estradiol replacement on brain-derived neurotrophic factor messenger ribonucleic acid expression in cortical and hippocampal brain regions of female Sprague–Dawley rats. Endocrinology 1995; 136: 2320–2324.
14. Bimonte-Nelson CA, Nelson ME, Granholm AC. Progesterone counteracts estrogen-induced increases in neurotrophins in the aged female rat brain. Neuroreport 2004; 15:2659–2663.
15. Ivanov T, Karolczak M, Beyer C. Estradiol stimulates GDNF expression in developing hypothalamic neurons. Endocrinology 2002; 143:3175–3178.
16. Zhang L, Chang YH, Barker JL, Hu Q, Maric D, Li BS, Rubinow DR. Testosterone and estrogen affect neuronal differentiation but not proliferation in early embryonic cortex of the rat: the possible roles of androgen and estrogen receptors. Neurosci Lett 2000; 231:57–60.
17. Kang JH, Lee CK, Kim JR, Yu SJ, Jo JH, Do BR, Kim HK, Kang SG.Estrogen stimulates the neuronal differentiation of human umbilical cord blood mesenchymal stem cells (CD34-). Neuroreport. 2007; 18: 35-38.
18. Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, Pell CL, Johnstone BH, Considine RV, March KL. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation 2004; 109: 1292–1298.
19. Okada M, Murase K, Makino A, Nakajima M, Kaku T, Furukawa S, Furukawa Y. Effects of estrogens on proliferation and differentiation of neural stem/progenitor cells. Biomed Res. 2008; 29:163-70.
20.  Razavi S, Razavi MR, Zarkesh Esfahani H, Kazemi M, Mostafavi FS Comparing brain-derived neurotrophic factor and ciliary neurotrophic factor secretion of induced neurotrophic factor secreting cells from human adipose and bone marrow-derived stem cells. Dev Growth Differ. 2013; 55: 648-55.
21.  Wei X, Zhao L, Zhong J, Gu H, Feng D, Johnstone BH, March KL, Farlow MR, Du Y. Adipose stromal cells-secreted neuroprotective media against neuronal apoptosis. Neurosci Lett.2009; 462: 76–79.
22. Razavi S, Mardani M, Kazemi M, Esfandiari E, Narimani M, Esmaeili A, Ahmadi N. Effect of leukemia inhibitory factor on the myelinogenic ability of Schwann-like cells induced from human adipose-derived stem cells.Cell Mol Neurobiol. 2013; 33: 283-289.
23. Razavi S, Razavi MR, Kheirollahi-Kouhestani M, Mardani M, Mostafavi FS. Co-culture with neurotrophic factor secreting cells induced from adipose-derived stem cells: promotes neurogenic differentiation.Biochem Biophys Res Commun. 2013 25; 440:381-387.
24. Su JD, Qiu J, Zhong YP, Li XY, Wang JW, Chen YZ. Expression of estrogen receptor (ER)-alpha and -beta immunoreactivity in hippocampal cell cultures with special attention to GABAergic neurons. J Neurosci Res 2001; 65:396-402.
25. Pilgrim C, Hutchison JB. Developmental regulation of sex differences in the brain: Can the role of gonadal steroids be redefined? Neuroscience 1994; 60: 843–855.
26. McEwen BS, Alves SE, Bulloch K, Weiland NG. Ovarian steroids and the brain: Implications for cognition and aging. Neurology 1997; 48: S8–15.
27. Lafferty FW, Fiske ME. Postmenopausal estrogen replacement: A long-term cohort study. Am J Med 1994; 97: 66–77.
28. Hurn PD, Macrae IM. Estrogen as a neuroprotectant in stroke. J Cereb Blood Flow Metab. 2000; 20: 631- 652.
29. Evans RM. The steroid and thyroid hormone receptor superfamily. Science 1988; 240: 889-895.
30. Paech K, Webb P, Kuiper GG, Nilsson S, Gustafsson J, Kushner PJ, Scanlan TS. Differential ligand activation of estrogen receptors ERα and ERβ at AP1 sites.Science1997; 277: 1508–1510.
31. Patrone C, Pollio G, Vegeto E, Enmark E, de Curtis I, Gustafsson JA, Maggi A. Estradiol induces differential neuronal phenotypes by activating estrogen receptor alpha or beta. Endocrinology 2000; 141: 1839–1845.
32. Nilsson S, Mäkelä S, Treuter E, Tujague M, Thomsen J, Andersson G, Enmark E, Pettersson K, Warner M, Gustafsson JA. Mechanisms of estrogen action. Physiol Rev 2001; 81: 1535– 1565.
33. Patrone C, Andersson S, Korhonen L, Lindholm D.  Estrogen receptor-dependent regulation of sensory neuron survival in developing dorsal root ganglion. Proc Natl Acad Sci U S A. 1999; 96: 10905–10910.
34. Almeida RD, Manadas BJ, Melo CV, Gomes JR, Mendes CS, Grãos MM, Carvalho RF, Carvalho AP, Duarte CB. Neuroprotection by BDNF against glutamate induced apoptotic cell death is mediated by ERK and PI3-kinase pathways. Cell Death Differ 2005; 12: 1329–1343.
35. Hong SH, Nah HY, Lee YJ, Lee JW, Park JH, Kim SJ, Lee JB, Yoon HS, Kim CH.Expression of estrogen receptor- alpha and –beta, glucocorticoid receptor, and progesterone receptor genes in human embryonic stem cells and embryoid bodies. Mol Cells 2004; 18: 320–325.
36. Wong JK, Le HH, Zsarnovszky A, Belcher SM. Estrogens and ICI182, 780 (Faslodex) modulate mitosis and cell death in immature cerebellar neurons via rapid activation of p44/p42 mitogen-activated protein kinase.J Neurosci. 2003; 23: 4984-4995.
37. Hamada H, Kim MK, Iwakura A, Ii M, Thorne T, Qin G, Asai J, Tsutsumi Y, Sekiguchi H, Silver M, Wecker A, Bord E, Zhu Y, Kishore R, Losordo DW. Estrogen receptors alpha and beta mediate contribution of bone marrow-derived endothelial progenitor cells to functional recovery after myocardial infarction. Circulation 2006, 114: 2261–2270.