Endometrial stem cells differentiation into neural cells by LY294002 small molecule

How to Cite:Homa Mohseni KouchesfahaniSomayeh Ebrahimi Baroughjafar AiHamideh AnbarEndometrial stem cells differentiation into neural cells by LY294002 small molecule.koomesh.18(1):e151146.

Abstract

Introduction: Endometrial stem cells (EnSCs) were identified for the first time in 2004. These cells are capable of extensive self-renewal and have the potency to differentiate into chondrocyte, osteocyte, adipocyte, neuron and oligodendrocyte. PI3K/Akt signaling has been implicated in multiple cellular and organ functions, including differentiation, survival and cell death and its inhibition leads to cell differentiation. The purpose of this study was to investigate the differentiation of endometrial stem cells into neural cells by inhibition of PI3K/Akt pathway using small molecule Ly294002. Materials and Methods: Endometrial tissues were treated enzymatically and segregated cells were cultured in DMEM/F12 with 10% FBS. The flow cytometry analysis was perfomed for CD105, CD90, CD146, CD31 and CD34 at the third passage. Then the neurogenic differentiation was evaluated at the third passage, 21 days after induction with differentiation media. Immunocytochemistry and Real-time PCR were performed to investigate the expression of specific neural stem cells markers. Results: The flow cytometry analysis showed that EnSCs were positive for CD90, CD105 and CD146 and negative for CD31 and CD34. Immunocytochemistry showed that the expression of nestin, NF and Chat neuronal markers in the cells treated with small molecule Ly294002. Real-time PCR also indicated expression of NF and Chat neuronal markers at the mRNA level. Conclusion: According to the findings of this study it can be concluded that the EnSCs have neural differentiation potency in the suitable differentiation milieu. Ly294002 small molecules by inhibiting PI3K / Akt pathway possibly can prevent cell proliferation and induce cell differentiation

Keywords

Phosphatidylinositol 3-Kinases

References

1.
Sakaki-Yumoto M, Katsuno Y, Derynck R. TGF-beta family signaling in stem cells. Biochim Biophys Acta 2013; 1830: 2280-96.
2.
Hombach-Klonisch S, Panigrahi S, Rashedi I, Seifert A, Alberti E, Pocar P, et al. Adult stem cells and their trans-differentiation potential--perspectives and therapeutic applications. J Mol Med (Berl) 2008; 86: 1301-1314.
3.
Lakshmipathy U, Verfaillie C. Stem cell plasticity. Blood reviews 2005; 19: 29-38.
4.
Ding S, Schultz PG. A role for chemistry in stem cell biology. Nature Biotechnol 2004; 22: 833-840.
5.
Henningson CT Jr, Stanislaus MA, Gewirtz AM. 28. Embryonic and adult stem cell therapy. J Allergy Clin Immunol 2003; 111: S745-53.
6.
Amoh Y, Katsuoka K, Hoffman RM. The advantages of hair follicle pluripotent stem cells over embryonic stem cells and induced pluripotent stem cells for regenerative medicine. J Dermatologi Sci 2010; 60: 131-137.
7.
Barry FP, Murphy JM. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol 2004; 36: 568-584.
8.
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143-147.
9.
Peister A, Mellad JA, Larson BL, Hall BM, Gibson LF, Prockop DJ. Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood 2004; 103: 1662-1668.
10.
Orbay H, Tobita M, Mizuno H. Mesenchymal stem cells isolated from adipose and other tissues: basic biological properties and clinical applications. Stem cells Int 2012; 2012: 461-718.
11.
Wolff EF, Wolff AB, Hongling D, Taylor HS. Demonstration of multipotent stem cells in the adult human endometrium by in vitro chondrogenesis. Reprod Sci 2007; 14: 524-533.
12.
Mobarakeh ZT, Ai J, Yazdani F, Sorkhabadi SM, Ghanbari Z, Javidan AN, et al. Human endometrial stem cells as a new source for programming to neural cells. Cell Biol Intern Report 2012; 19: e00015.
13.
Ebrahimi-Barough S, Kouchesfahani HM, Ai J, Massumi M. Differentiation of human endometrial stromal cells into oligodendrocyte progenitor cells (OPCs). J Mol Neurosci 2013; 51: 265-273.
14.
Ai J, Shahverdi AR, Barough SE, Kouchesfehani HM, Heidari S, Roozafzoon R, Verdi J, Khoshzaban A. Derivation of adipocytes from human endometrial stem cells (EnSCs). J Reprod Infertil 2012; 13: 151-157.
15.
Wang Y, He W, Bian H, Liu C, Li S. Small molecule induction of neural-like cells from bone marrow-mesenchymal stem cells. J Cell Biochem 2012; 113: 1527-1536.
16.
Gharbi SI, Zvelebil MJ, Shuttleworth SJ, Hancox T, Saghir N, Timms JF, Waterfield MD. Exploring the specificity of the PI3K family inhibitor LY294002. Biochem J 2007; 404: 15-21.
17.
Li W, Ding S. Small molecules that modulate embryonic stem cell fate and somatic cell reprogramming. Trends Pharmacol Sci 2010; 31: 36-45.
18.
Cantley LC. The phosphoinositide 3-kinase pathway. Science 2002; 296: 1655-1657.
19.
Wu B, Li W, Wang L, Liu ZH, Zhao XY. Stem cells and small molecule screening: haploid embryonic stem cells as a new tool. Acta Pharmacol Sin 2013; 34: 725-731.
20.
Schugar RC, Robbins PD, Deasy BM. Small molecules in stem cell self-renewal and differentiation. Gene Ther 2008; 15: 126-135.
21.
Faghihi F, Baghaban Eslaminejad M, Nekookar A, Najar M, Salekdeh GH. The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. Biomed Pharmacother 2013; 67: 31-38.
22.
Lu B, Atala A. Small molecules and small molecule drugs in regenerative medicine. Drug Discov today 2014; 19: 801-808.
23.
Song H, Chang W, Song BW, Hwang KC. Specific differentiation of mesenchymal stem cells by small molecules. Am J Stem Cells 2012; 1: 22-30.
24.
Vanhaesebroeck B, Stephens L, Hawkins P. PI3K signalling: the path to discovery and understanding. Nature Rev Mol Cell Biol 2012; 13: 195-203.
25.
Foster FM, Traer CJ, Abraham SM, Fry MJ. The phosphoinositide (PI) 3-kinase family. J Cell Sci 2003; 116: 3037-3040.
26.
Hemmings BA, Restuccia DF. PI3K-PKB/Akt pathway. Cold Spring Har Perspect Biol 2012; 4: a011189.
27.
Woodbury D, Schwarz EJ, Prockop DJ, Black IB. Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 2000; 61: 364-370.
28.
Taha MF, Javeri A, Kheirkhah O, Majidizadeh T, Khalatbary AR. Neural differentiation of mouse embryonic and mesenchymal stem cells in a simple medium containing synthetic serum replacement. J Biotechnol 2014; 172: 1-10.
29.
Choi YK, Cho H, Seo YK, Yoon HH, Park JK. Stimulation of sub-sonic vibration promotes the differentiation of adipose tissue-derived mesenchymal stem cells into neural cells. Life Sci 2012; 91: 329-337.
30.
Yan ZJ, Hu YQ, Zhang HT, Zhang P, Xiao ZY, Sun XL, et al. Comparison of the neural differentiation potential of human mesenchymal stem cells from amniotic fluid and adult bone marrow. Cell Mol Neurobiol 2013; 33: 465-475.
31.
Zhao P, Ise H, Hongo M, Ota M, Konishi I, Nikaido T. Human amniotic mesenchymal cells have some characteristics of cardiomyocytes. Transplantation 2005; 79: 528-535.
32.
Kim EY, Lee KB, Kim MK. The potential of mesenchymal stem cells derived from amniotic membrane and amniotic fluid for neuronal regenerative therapy. BMB Report 2014; 47: 135-140.
33.
Kadivar M, Piryaei F, Ramezani M. Comparison of the differentiation potential of human mesenchymal stem cells and several animal species. Koomesh 2010; 11: 270-278. (Persian).
34.
Hoveizi E, Nabiuni M, Massumi M, Parivar K. Derivation of definitive endoderm from human induced pluripotent stem cells using signaling molecules. Koomesh 2014; 15: 212-219. (Persian).
35.
Wolff EF, Gao XB, Yao KV, Andrews ZB, Du H, Elsworth JD, Taylor HS. Endometrial stem cell transplantation restores dopamine production in a Parkinson's disease model. J Cell Mol Med 2011; 15: 747-755.
36.
Ebrahimi-Barough S, Norouzi Javidan A, Saberi H, Joghataei MT, Rahbarghazi R, Mirzaei E, et al. Evaluation of motor neuron-Like cell differentiation of hEnSCs on biodegradable PLGA nanofiber scaffolds. Mol Neurobiol 2015; 52: 1704-1713.
37.
Navaei-Nigjeh M, Amoabedini G, Noroozi A, Azami M, Asmani MN, Ebrahimi-Barough S, et al. Enhancing neuronal growth from human endometrial stem cells derived neuron-like cells in three-dimensional fibrin gel for nerve tissue engineering. J Biomed Mater Res A 2014; 102: 2533-2543.

comments