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Cytotechnology

, Volume 66, Issue 1, pp 119–131 | Cite as

The simulated microgravity enhances multipotential differentiation capacity of bone marrow mesenchymal stem cells

  • Nanding Wang
  • Huan Wang
  • Jun Chen
  • Xiaofeng Zhang
  • Juan Xie
  • Zhi Li
  • Jing Ma
  • Wen Wang
  • Zongren Wang
Original Research

Abstract

Multi-differentiation capability is an essential characteristic of bone marrow mesenchymal stem cells (BMSCs). Method on obtaining higher-quality stem cells with an improved differentiation potential has gained significant attention for the treatment of clinical diseases and developmental biology. In our study, we investigated the multipotential differentiation capacity of BMSCs under simulated microgravity (SMG) condition. F-actin staining found that cytoskeleton took on a time-dependent change under SMG condition, which caused spindle to round morphological change of the cultured cells. Quantitative PCR and Western Blotting showed the pluripotency marker OCT4 was up-regulated in the SMG condition especially after SMG of 72 h, which we observed would be the most appropriate SMG duration for enhancing pluripotency of BMSCs. After dividing BMSCs into normal gravity (NG) group and SMG group, we induced them respectively in endothelium oriented, adipogenic and neuronal induction media. Immunostaining and Western Blotting found that endothelium oriented differentiated BMSCs expressed higher VWF and CD31 in the SMG group than in the NG group. The neuron-like cells derived from BMSCs in the SMG group also expressed higher level of MAP2 and NF-H. Furthermore, the quantity of induced adipocytes increased in the SMG group compared to the NG group shown by Oil Red O staining, The expression of PPARγ2 increased significantly under SMG condition. Therefore, we demonstrated that SMG could promote BMSCs to differentiate into many kinds of cells and predicted that enhanced multi-potential differentiation capacity response in BMSCs following SMG might be relevant to the changes of cytoskeleton and the stem cell marker OCT4.

Keywords

Simulated microgravity Bone marrow mesenchymal stem cells Pluripotency Differentiation OCT4 

Abbreviations

SMG

Simulated microgravity

NG

Normal gravity

MSC

Mesenchymal stem cell

BMSC

Bone marrow mesenchymal stem cell

VWF

Von Willebrand factor

MAP2

Microtubule-associated protein 2

NF-H

Neurofilament heavy chain

OCT4

Octamer-binding transcription factor 4

Notes

Acknowledgments

This work was carried out in the Physiology laboratory and State Key Laboratory of Aerospace Biodynamics at Fourth Military Medical University. The project was supported by NSFC Grant 30973808.

Supplementary material

10616_2013_9544_MOESM1_ESM.tif (16.6 mb)
Supplementary material 1 (TIFF 16991 kb)

References

  1. Barry FP, Murphy JM (2004) Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol 36:568–584CrossRefGoogle Scholar
  2. Basso N, Bellows CG, Heersche JN (2005) Effect of simulated weightlessness on osteoprogenitor cell number and proliferation in young and adult rats. Bone 36:173–183CrossRefGoogle Scholar
  3. Chen J, Liu R, Yang Y, Li J, Zhang X, Li J, Wang Z, Ma J (2011) The simulated microgravity enhances the differentiation of mesenchymal stem cells into neurons. Neurosci Lett 505:171–175CrossRefGoogle Scholar
  4. Colter DC, Class R, DiGirolamo CM, Prockop DJ (2000) Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci USA 97:3213–3218CrossRefGoogle Scholar
  5. Crisostomo PR, Wang M, Wairiuko GM, Morrell ED, Terrell AM, Seshadri P, Nam UH, Meldrum DR (2006) High passage number of stem cells adversely affects stem cell activation and myocardial protection. Shock 26:575–580CrossRefGoogle Scholar
  6. Gershkovich PM, Gershkovich I, Buravkova LB (2011) Expression of cytoskeleton genes in culture of human mesenchymal stromal cells in different periods of simulating the effects of microgravity. Aviakosm Ekolog Med 45(4):39–41Google Scholar
  7. Graziano A, D’Aquino R, Cusella-De AM, Laino G, Piattelli A, Pacifici M, De Rosa A, Papaccio G (2007) Concave pit-containing scaffold surfaces improve stem cell-derived osteoblast performance and lead to significant bone tissue formation. PLoS One 2:e496CrossRefGoogle Scholar
  8. Hosu BG, Mullen SF, Critser JK, Forgacs G (2008) Reversible disassembly of the actin cytoskeleton improves the survival rate and developmental competence of cryopreserved mouse oocytes. PLoS One 3:e2787CrossRefGoogle Scholar
  9. Jin Y, Liu Y, Antonyak M, Peng X (2012) Isolation and characterization of vascular endothelial cells from murine heart and lung. Methods Mol Biol 843:147–154CrossRefGoogle Scholar
  10. Kasper G, Dankert N, Tuischer J, Hoeft M, Gaber T, Glaeser JD, Zander D, Tschirschmann M, Thompson M, Matziolis G, Duda GN (2007) Mesenchymal stem cells regulate angiogenesis according to their mechanical environment. Stem Cells 25:903–910CrossRefGoogle Scholar
  11. Kilian KA, Bugarija B, Lahn BT, Mrksich M (2010) Geometric cues for directing the differentiation of mesenchymal stem cells. Proc Natl Acad Sci USA 107:4872–4877CrossRefGoogle Scholar
  12. Klaus DM (2001) Clinostats and bioreactors. Gravit Space Biol Bull 14:55–64Google Scholar
  13. Kodama H, Inoue T, Watanabe R, Yasuoka H, Kawakami Y, Ogawa S, Ikeda Y, Mikoshiba K, Kuwana M (2005) Cardiomyogenic potential of mesenchymal progenitors derived from human circulating CD14+ monocytes. Stem Cells Dev 14:676–686CrossRefGoogle Scholar
  14. Kodama H, Inoue T, Watanabe R, Yasutomi D, Kawakami Y, Ogawa S, Mikoshiba K, Ikeda Y, Kuwana M (2006) Neurogenic potential of progenitors derived from human circulating CD14+ monocytes. Immunol Cell Biol 84:209–217CrossRefGoogle Scholar
  15. Koike M, Shimokawa H, Kanno Z, Ohya K, Soma K (2005) Effects of mechanical strain on proliferation and differentiation of bone marrow stromal cell line ST2. J Bone Miner Metab 23:219–225CrossRefGoogle Scholar
  16. Li J, Zhang S, Chen J, Du T, Wang Y, Wang Z (2009) Modeled microgravity causes changes in the cytoskeleton and focal adhesions, and decreases in migration in malignant human MCF-7 cells. Protoplasma 238:23–33CrossRefGoogle Scholar
  17. Mauney JR, Sjostorm S, Blumberg J, Horan R, O’Leary JP, Vunjak-Novakovic G, Volloch V, Kaplan DL (2004) Mechanical stimulation promotes osteogenic differentiation of human bone marrow stromal cells on 3-d partially demineralized bone scaffolds in vitro. Calcif Tissue Int 74:458–468CrossRefGoogle Scholar
  18. McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS (2004) Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 6:483–495CrossRefGoogle Scholar
  19. McBride SH, Falls T, Knothe TM (2008) Modulation of stem cell shape and fate b: mechanical modulation of cell shape and gene expression. Tissue Eng Part A 14:1573–1580CrossRefGoogle Scholar
  20. Muruganandan S, Parlee SD, Rourke JL, Ernst MC, Goralski KB, Sinal CJ (2011) Chemerin, a novel peroxisome proliferator-activated receptor gamma (PPARgamma) target gene that promotes mesenchymal stem cell adipogenesis. J Biol Chem 286:23982–23995CrossRefGoogle Scholar
  21. Niwa H, Miyazaki J, Smith AG (2000) Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat Genet 24:372–376CrossRefGoogle Scholar
  22. Oswald J, Boxberger S, Jorgensen B, Feldmann S, Ehninger G, Bornhauser M, Werner C (2004) Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells 22:377–384CrossRefGoogle Scholar
  23. Owen M, Friedenstein AJ (1988) Stromal stem cells: marrow-derived osteogenic precursors. Ciba Found Symp 136:42–60Google Scholar
  24. Phinney DG, Prockop DJ (2007) Concise review: mesenchymal stem/multipotent stromal cells: the state of transdifferentiation and modes of tissue repair—current views. Stem Cells 25:2896–2902CrossRefGoogle Scholar
  25. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147CrossRefGoogle Scholar
  26. Prockop DJ (1997) Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74CrossRefGoogle Scholar
  27. Ruggeri ZM (2003) Von Willebrand factor, platelets and endothelial cell interactions. J Thromb Haemost 1:1335–1342CrossRefGoogle Scholar
  28. Tamama K, Fan VH, Griffith LG, Blair HC, Wells A (2006) Epidermal growth factor as a candidate for ex vivo expansion of bone marrow-derived mesenchymal stem cells. Stem Cells 24:686–695CrossRefGoogle Scholar
  29. Tamama K, Sen CK, Wells A (2008) Differentiation of bone marrow mesenchymal stem cells into the smooth muscle lineage by blocking ERK/MAPK signaling pathway. Stem Cells Dev 17:897–908CrossRefGoogle Scholar
  30. Xiang Y, Zheng Q, Jia B, Huang G, Xie C, Pan J, Wang J (2007) Ex vivo expansion, adipogenesis and neurogenesis of cryopreserved human bone marrow mesenchymal stem cells. Cell Biol Int 31:444–450CrossRefGoogle Scholar
  31. Yoshikawa T, Peel SA, Gladstone JR, Davies JE (1997) Biochemical analysis of the response in rat bone marrow cell cultures to mechanical stimulation. Biomed Mater Eng 7:369–377Google Scholar
  32. Yuge L, Hide I, Kumagai T, Kumei Y, Takeda S, Kanno M, Sugiyama M, Kataoka K (2003) Cell differentiation and p38(MAPK) cascade are inhibited in human osteoblasts cultured in a three-dimensional clinostat. In Vitro Cell Dev Biol Anim 39:89–97CrossRefGoogle Scholar
  33. Yuge L, Kajiume T, Tahara H, Kawahara Y, Umeda C, Yoshimoto R, Wu SL, Yamaoka K, Asashima M, Kataoka K, Ide T (2006) Microgravity potentiates stem cell proliferation while sustaining the capability of differentiation. Stem Cells Dev 15:921–929CrossRefGoogle Scholar
  34. Yuge L, Sasaki A, Kawahara Y, Wu SL, Matsumoto M, Manabe T, Kajiume T, Takeda M, Magaki T, Takahashi T, Kurisu K, Matsumoto M (2011) Simulated microgravity maintains the undifferentiated state and enhances the neural repair potential of bone marrow stromal cells. Stem Cells Dev 20:893–900CrossRefGoogle Scholar
  35. Zhang X, Nan Y, Wang H, Chen J, Wang N, Xie J, Ma J, Wang Z (2013) Model microgravity enhances endothelium differentiation of mesenchymal stem cells. Naturwissenschaften 100:125–133CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Nanding Wang
    • 1
  • Huan Wang
    • 2
  • Jun Chen
    • 3
  • Xiaofeng Zhang
    • 1
  • Juan Xie
    • 1
  • Zhi Li
    • 4
  • Jing Ma
    • 1
  • Wen Wang
    • 1
  • Zongren Wang
    • 1
  1. 1.Department of Traditional Chinese Medicine, Xijing HospitalFourth Military Medical UniversityXi’anPeople’s Republic of China
  2. 2.Department of Dermatology, Tangdu HospitalFourth Military Medical UniversityXi’anPeople’s Republic of China
  3. 3.Department of EncephalopathyTraditional Chinese Medicine Hospital of Shaanxi ProvinceXi’anPeople’s Republic of China
  4. 4.Department of Cardiovascular Surgery, Xijing HospitalFourth Military Medical UniversityXi’anPeople’s Republic of China

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