Expression pattern of neurotrophins and their receptors during neuronal differentiation of adipose-derived stem cells in simulated microgravity condition

Document Type : Original Article

Authors

1 Department of Biology, Fars Science and Research Branch, Islamic Azad University, Marvdasht, Iran

2 Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran

3 Aerospace Research Institute, Ministry of Science Research and Technology, Tehran, Iran

Abstract

Objective(s): Studies have confirmed that microgravity, as a mechanical factor, influences both differentiation and function of mesenchymal stem cells. Here we investigated the effects of simulated microgravity on neural differentiation of human adipose-derived stem cells (ADSCs).
Materials and Methods:We have used a fast rotating clinostat (clinorotation) to simulate microgravity condition. Real-time PCR and flow cytometry analysis were used to evaluate the regulation of neurotrophins, their receptors, and neural markers by simulated microgravity and their impact on neural differentiation of cells.
Results: Our data revealed that simulation microgravity up-regulated the expression of MAP-2, BDNF, TrkB, NT-3, and TrkC both before and after neural differentiation. Also, the neural cells derived from ADSCs in microgravity condition expressed more MAP-2, GFAP, and synaptophysin protein in comparison to the 1G control.
Conclusion: We showed that simulated microgravity can enhance the differentiation of mesenchymal stem cells into neurons. Our findings provide a new strategy for differentiation of ADSCs to neural-like cells and probably other cell lineages. Meanwhile, microgravity simulation had no adverse effects on the viability of the cells and could be used as a new environment to successfully manipulate cells.

Keywords

References

1.Arvidsson A, Collin T, Kirik D, Kokai Z, Lindvall O. Neuronal replacement from endogenous precursors in the adult brain after stroke. Nat Med 2002; 8:963–970.
2. Singec I, Jandial R, Crain A, Nikkhah G, Snyder EY. The leading edge of stem cell therapeutics. Annu Rev Med 2007; 58:313–328.
3.Zietlow R, Lane EL, Dunnett SB, Rosser AE. Human stem cells for CNS repair. Cell Tissue Res 2008; 331:301–322.
4.Ninomiya M, Yamashita T, Araki N, Okano H, Sawamoto K. Enhanced neurogenesis in the ischemic striatum following EGF-induced expansion of transit-amplifying cells in the subventricular zone. Neurosci Lett 2006; 403:63–67.
5.Aharonowiz M, Einstein O, Fainstein N, Lassmann H, Reubinoff B, Ben-Hur T. Neuroprotective effect of transplanted human embryonic stem cell-derived neural precursors in an animal model of multiple sclerosis. PLoS One 2008; 3:e3145.
6.Einstein O, Grigoriadis N, Mizrachi-Kol R, Reinhartz E, Polyzoidou E, Lavon I. Transplanted neural precursor cells reduce brain inflammation to attenuate chronic experimental autoimmune encephalomyelitis. Exp Neurol 2006; 198:275-284.
7.Hellmann M, Panet H, Barhum Y, Melamed E. D Offen. Increased survival and migration of engrafted mesenchymal bone marrow stem cells in 6-hydroxydopamine-lesioned rodents. Neurosci Lett 2006; 395:124-128.
8.Cuevas P, Carceller F, Dujovny M, Garcia-Gomez I, Cuevas B, Gonzalez-Corrochano R, et al. Peripheral nerve regeneration by bone marrow stromal cells Neurol Res 2002; 24:634–638.
9.Mahmood A, Lu D, Lu M, Chopp M. Treatment of traumatic brain injury in adult rats with intravenous administration of human bone marrow stromal cells. Neurosurgery 2003; 53:697–702.
10.Bradamante S, Barenghi L, Maier M. Stem cells toward the future: The space challenge. Life 2014; 4:267-280.
11.Zuk P, Zhu M, Mizuno H, Huang J, Futrell J, Katz A, et al. Multilineage Cells from Human Adipose Tissue: Implications for Cell-Based Therapies. Tissue Engineering 2001; 7:211-228.
12.Erices A, Conget P, Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol 2000; 109: 235-242.
13.Roufosse CA, Direkze,NC, Otto WR, Wright NA. Circulating mesenchymal stem cells. Int J Biochem Cell Biol 2004; 36: 585-597.
14.Haniffa M, Wang X, Holtick U, Rae M, Isaacs J, Dickinson A, et al. Adult human Fibroblasts Are Potent Immunoregulatory Cells and Functionally Equivalent to mesenchymal stem cells. J Immunol 2007; 179:1595-1604.
15.Sessarego N, Parodi A, Podesta M, Benvenuto F, Mogni M, Raviolo, V, et al. Multipotent mesenchymal stromal cells from amniotic fluid: solid perspectives for clinical application. Haematologica 2008; 93:339-346.
16.Yan X, Fu C, Chen L, Qin J, Zeng Q, Yuan H, et al. Mesenchymal stem cells from primary breast cancer tissue promote cancer proliferation and enhance mammosphere formation partially via EGF/EGFR/Akt pathway. Breast Cancer Res Treat 2011; 132: 153-164.
17.Birmingham E, Niebur GL, McHugh PE, Shaw G, Barry FP, McNamara LM. Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche. Eur Cell Mater 2012; 23:13-27.
18.Fink T, Zachar V. Adipogenic differentiation of human mesenchymal stem cells. Methods Mol Biol 2011; 698:243-251.
19.Alves da Silva ML, Martins A, Costa-Pinto AR, Correlo VM, Sol P,  Bhattacharya M, et al. Chondrogenic differentiation of human bone marrow mesenchymal stem cells in chitosan-based scaffolds using a flow-perfusion bioreactor. J Tissue Eng Regen Med 2011; 5:722–732.
20.Carvalho PH, Daibert AP, Monteiro BS, Okano BS, Carvalho JL, Cunha DN, et al. Differentiation of adipose tissue-derived mesenchymal stem cells into cardiomyocytes. Arq Bras Cardiol 2013; 100:82-89.
21.Lee CH, Moioli EK, Mao JJ. Fibroblastic differentiation of human mesenchymal stem cells using connective tissue growth factor. Conf Proc IEEE Eng Med Biol Soc 2006; 1:775-778.
22.Chowdhury R, Webber JP, Gurney M, Mason MD, Tabi Z, Clayton A. Cancer exosomes trigger mesenchymal stem cell differentiation into pro-angiogenic and pro-invasive myofibroblasts. Oncotarget 2015; 20:715-731.
23.Păunescu V, Deak E, Herman D,  Siska I R, T˘anasie G, Bunu C, et al. In vitro differentiation of human mesenchymal stem cells to epithelial lineage. J Cell Mol Med 2007; 11: 502–508.
24.Wu XB, Tao R. Hepatocyte differentiation of mesenchymal stem cells. Hepatobiliary Pancreat Dis Int 2012; 11:360-371.
25.Woodbury D, Schwarz EJ, Prockop DJ, Black IB. Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 2000; 15; 61:364-370.
26.Hofstetter C, Schwartz E, Hess D, Widenfalk J, El Manira A, Prockop D J, et al. Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc Natl Acad Sci USA 2002; 99:2199–2204.
27.Lu P, Jones L, Tuszynski M. BDNF-expressing marrow stromal cells support extensive axonal growth at sites of spinal cord injury. Exp Neurol 2005; 191: 344–360.
28.Mahmood A, Lu D, Lu M, Chopp M. Treatment of traumatic brain injury in adult rats with intravenous administration of human bone marrow stromal cells. Neurosurgery 2003; 53:697–702.
29.Guo L, Yin F, Meng HQ, Ling L, Huhe T, Li P, et al. Differentiation of mesenchymal stem cells into dopaminergic neuron-like cells in vitro. Biomed  environ sci 2005; 18:36-42.
30.Shinde V, Brungs S, Henry M, Wegener L, Nemade H, Rotshteyn T, et al. Simulated microgravity modulates differentiation processes of embryonic stem cells. Cell Physiol Biochem 2016; 38:1483-1499.
31.Blaber E, Sato K, Almeida E. Stem cell health and tissue regeneration in microgravity. Stem Cells Dev 2014; 23:73-78.
32.Silvano M, Miele E, Valerio M, Casadei L, Begalli F, Campese A, et al. Consequences of simulated microgravity in neural stem cells: Biological effects and metabolic response. J Stem Cell Res Ther 2015; 5:289-296.
33.Hart DA. Is Adipocyte differentiation the default lineage for mesenchymal stem/progenitor cells after loss of mechanical loading? A perspective from space flight and model systems.  J Biomed Sci Eng 2014; 7:799-808.
34.Grimm D, Bauer J, Kossmehl P, Shakibaei M, Schönberger J, Pickenhahn H, et al. Simulated microgravity alters differentiation ad increases apoptosis in human follicular thyroid carcinoma cells. FASEB J 2002; 16:604-614.
35.Claassen DE, Spooner BS. Impact of altered gravity on aspects of cell biology. Int Rev Cyt 1994; 156: 301– 373.
36.Hammond TG, Lewis FC, Goodwin TJ, Linnehan RM, Wolf DA, Hire KP, et al. Gene expression in space. Nat Med 1999; 5: 359.
37.Unsworth BR, Lelkes PI. Growing tissues in microgravity. Nat Med 1998; 4:901-907.
38.Zuk PA, Zhu M, Mizuno H, Huang JI, Futrell WJ, Katz AJ, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001; 7:211–226.
39.Mitchell JB, McIntosh K, Zvonic S. Immunophenotype of human adipose-derived cells: Temporal changes in stromalassociated and stem cell-associated markers. Stem Cells 2006; 24:376.
40.Chen JY, Mou XZ, Du XC, Xiang C. Comparative analysis of biological characteristics of adult mesenchymal stem cells with different tissue origins. Asian Pac J Trop Med 2015; 8:739–746.
41.Edalat H, Hajebrahimi Z, Movahedin M, Tavallaei M, Amiri S, Mowlaa S J. p75NTR suppression in rat bone marrow stromal stem cells significantly reduced their rate of apoptosis during neural differentiation. Neurosci Lett 2011; 498 :15– 19.
42.Dominici M, Le Blanc K, Mueller I, Slaper- Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement, Cytotherapy 2006; 8:315-317.
43.Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A. Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol 2000; 164: 247–256.
44.Tondreau T, Lagneaux L, Dejeneffe M, Massy M, Mortier C, Delforge A. Bone marrow-derived mesenchymal stem cells already express specific neural proteins before any differentiation. Differentiation 2004; 72: 319–326.
45.Hermann A, Maisel M, Storch A. Epigenetic conversion of human adult bone mesodermal stromal cells into neuroectodermal cell types for replacement therapy of neurodegenerative disorders. Expert Opin Biol Ther 2006; 6:653–670.
46.Greco SJ, Zhou C, Ye JH, Rameshwar P. An interdisciplinary approach and characterization of neuronal cells transdifferentiated from human mesenchymal stem cells. Stem Cells Dev 2007; 16:811–826.
47.Greco SJ, Zhou C, Ye JH, Rameshwar P. A method to generate human mesenchymal stem cell derived neurons which express and are excited by multiple neurotransmitters. Biol Proced Online 2008; 10:90–101.
48.Kan I, Ben-Zur T, Barhum Y, Levy YS, Burstein A, Charlow T, et al. Dopaminergic differentiation of human mesenchymal stem cells—utilization of bioassay for tyrosine hydroxylase expression. Neurosci Lett 2007; 419:28–33.
49.Barzilay R, Kan I, Ben-Zur T, Bulvik S, Melamed E, Offen D. Induction of human mesenchymal stem cells into dopamine-producing cells with different differentiation protocols. Stem Cells Dev 2008; 17:547–554.
50.Tondreau T, Dejeneffe M, Meuleman N, Stamatopoulos B, Delforge A, Martiat P. Gene expression pattern of functional neuronal cells derived from human bone marrow mesenchymal stromal cells. BMC Genomics 2008; 9: 166.
51.Chen J, Liu R, Yang Y, Li J, Zhang X, Li J, et al. The simulated microgravity enhances the differentiation of mesenchymal stem cells into neurons. Neuroscience Letters 2011; 505:171-175.
52.Yuge A, Sasaki Y, Kawahara SL, Wu M, Matsumoto T, Manabe T, et al. Simulated microgravity maintains the undifferentiated state and enhances the neural repair potential of bone marrow stromal cells. Stem Cells Dev 2010; 20: 893–900.
53.Rabizadeh S, Oh J, Zhong LT, Yang J, Bitler C M, Bredesen D E. Induction of apoptosis by the low-affinity NGF receptor. Science 1993; 261:345–348.
54.McBeath R, Pirone DM. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 2004; 6:483–495.
Iranian Journal of Basic Medical Sciences
Volume 20, Issue 2
February 2017
Pages 178-186

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