Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1
5-Year Impact Factor – 2.2
Scopus CiteScore – 3.4 (CiteScore Tracker 3.4)
Index Copernicus  – 161.11; MEiN – 140 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

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Advances in Clinical and Experimental Medicine

2018, vol. 27, nr 7, July, p. 971–977

doi: 10.17219/acem/70789

Publication type: original article

Language: English

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Evaluation of the secretion and release of vascular endothelial growth factor from two-dimensional culture and three-dimensional cell spheroids formed with stem cells and osteoprecursor cells

Hyunjin Lee1,A,B,C,D,E,F, Sung-Il Lee1,A,B,C,D,E,F, Youngkyung Ko1,A,B,C,D,E,F, Jun-Beom Park1,A,B,C,D,E,F

1 Department of Periodontics, College of Medicine, Catholic University of Korea, Seoul, the Republic of Korea

Abstract

Background. Co-culture has been applied in cell therapy, including stem cells, and has been reported to give enhanced functionality.
Objectives. In this study, stem-cell spheroids were formed in concave micromolds at different ratios of stem cells to osteoprecursor cells, and the amount of secretion of vascular endothelial growth factor (VEGF) was evaluated.
Material and Methods. Gingiva-derived stem cells and osteoprecursor cells in the amount of 6 × 105 were seeded on a 24-well culture plate or concave micromolds. The ratios of stem cells to osteoprecursor cells included: 0:4 (group 1), 1:3 (group 2), 2:2 (group 3), 3:1 (group 4), and 4:0 (group 5).
Results. The morphology of cells in a 2-dimensional culture (groups 1–5) showed a fibroblast-like appearance. The secretion of VEGF increased with the increase in stem cells, and a statistically significant increase was noted in groups 3, 4 and 5 when compared with the media-only group (p < 0.05). Osteoprecursor cells formed spheroids in concave microwells, and no noticeable change in the morphology was noted with the increase in stem cells. Spheroids containing stem cells were positive for the stem-cell markers SSEA-4. The secretion of VEGF from cell spheroids increased with the increase in stem cells.
Conclusion. This study showed that cell spheroids formed with stem cells and osteoprecursor cells with different ratios, using microwells, had paracrine effects on the stem cells. The secretion of VEGF increased with the increase in stem cells. This stem-cell spheroid may be applied for tissue-engineering purposes.

Key words

vascular endothelial growth factor, osteoblast, co-culture techniques, cellular spheroids, stem cell research

References (29)

  1. Wong SF, No da Y, Choi YY, Kim DS, Chung BG, Lee SH. Concave microwell based size-controllable hepatosphere as a three-dimensional liver tissue model. Biomaterials. 2011;32:8087–8096.
  2. Jun Y, Kang AR, Lee JS, et al. 3D co-culturing model of primary pancreatic islets and hepatocytes in hybrid spheroid to overcome pancreatic cell shortage. Biomaterials. 2013;34:3784–3794.
  3. Jin SH, Lee JE, Yun JH, Kim I, Ko Y, Park JB. Isolation and characterization of human mesenchymal stem cells from gingival connective tissue. J Periodontal Res. 2015;50:461–467.
  4. Jin SH, Kweon H, Park JB, Kim CH. The effects of tetracycline-loaded silk fibroin membrane on proliferation and osteogenic potential of mesenchymal stem cells. J Surg Res. 2014;192:1–9.
  5. Baraniak PR, McDevitt TC. Stem cell paracrine actions and tissue regeneration. Regen Med. 2010;5:121–143.
  6. Chan JK, Lam P. Soluble factors from human fetal bone marrow-derived mesenchymal stem cells: Preparation of conditioned medium and its effect on tumor cells. Methods Mol Biol. 2016;1416:467–475.
  7. Liang X, Ding Y, Zhang Y, Tse HF, Lian Q. Paracrine mechanisms of mesenchymal stem cell-based therapy: Current status and perspectives. Cell Transplant. 2014;23:1045–1059.
  8. Tang Y, Li Q, Meng F, et al. Therapeutic potential of HGF-expressing human umbilical cord mesenchymal stem cells in mice with acute liver failure. Int J Hepatol. 2016;2016:5452487.
  9. Yang Y, Hu S, Xu X, et al. The vascular endothelial growth factors-expressing character of mesenchymal stem cells plays a positive role in treatment of acute lung injury in vivo. Mediators Inflamm. 2016;2016:2347938.
  10. Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA. Vascular endothelial growth factor and angiogenesis. Pharmacol Rev. 2004;56(4):549–580.
  11. Kinnaird T, Stabile E, Burnett MS, et al. Local delivery of marrow-derived stromal cells augments collateral perfusion through paracrine mechanisms. Circulation. 2004;109:1543–1549.
  12. Shim EK, Lee JS, Kim DE, et al. Autogenous mesenchymal stem cells from the vertebral body enhance intervertebral disc regeneration by paracrine interaction: An in vitro pilot study. Cell Transplant. 2016; 25(10):1819–1832.
  13. Gnecchi M, Zhang Z, Ni A, Dzau VJ. Paracrine mechanisms in adult stem cell signaling and therapy. Circ Res. 2008;103:1204–1219.
  14. Geranmayeh MH, Baghbanzadeh A, Barin A, et al. Paracrine neuroprotective effects of neural stem cells on glutamate-induced cortical neuronal cell excitotoxicity. Adv Pharm Bull. 2015;5:515–521.
  15. Gokce A, Peak TC, Abdel-Mageed AB, Hellstrom WJ. Adipose tissue-derived stem cells for the treatment of erectile dysfunction. Curr Urol Rep. 2016;17:14.
  16. Izumoto-Akita T, Tsunekawa S, Yamamoto A, et al. Secreted factors from dental pulp stem cells improve glucose intolerance in streptozotocin-induced diabetic mice by increasing pancreatic beta-cell function. BMJ Open Diabetes Res Care. 2015;3:e000128.
  17. Lee SI, Yeo SI, Kim BB, Ko Y, Park JB. Formation of size-controllable spheroids using gingiva-derived stem cells and concave microwells: Morphology and viability tests. Biomed Rep. 2016;4:97–101.
  18. Gu Q, Tomaskovic-Crook E, Lozano R, et al. Stem cell bioprinting: Functional 3D neural mini-tissues from printed gel-based bioink and human neural stem cells. Adv Healthc Mater. 2016;5(12):1428.
  19. Seo S, Na K. Mesenchymal stem cell-based tissue engineering for chondrogenesis. J Biomed Biotechnol. 2011;2011:806891.
  20. Valarmathi MT, Yost MJ, Goodwin RL, Potts JD. The influence of proepicardial cells on the osteogenic potential of marrow stromal cells in a three-dimensional tubular scaffold. Biomaterials. 2008;29:2203–2216.
  21. Xu Y, Shi T, Xu A, Zhang L. 3D spheroid culture enhances survival and therapeutic capacities of MSCs injected into ischemic kidney. J Cell Mol Med. 2016;20(7):1203–1213.
  22. Zhu HX, Gao JL, Zhao MM, et al. Effects of bone marrow-derived mesenchymal stem cells on the autophagic activity of alveolar macrophages in a rat model of silicosis. Exp Ther Med. 2016;11:2577–2582.
  23. Sheashaa H, Lotfy A, Elhusseini F, et al. Protective effect of adipose-derived mesenchymal stem cells against acute kidney injury induced by ischemia-reperfusion in Sprague-Dawley rats. Exp Ther Med. 2016;11:1573–1580.
  24. Cui B, Li E, Yang B, Wang B. Human umbilical cord blood-derived mesenchymal stem cell transplantation for the treatment of spinal cord injury. Exp Ther Med. 2014;7:1233–1236.
  25. Fu Q, Tang NN, Zhang Q, et al. Preclinical study of cell therapy for osteonecrosis of the femoral head with allogenic peripheral blood-derived mesenchymal stem cells. Yonsei Med J. 2016;57:1006–1015.
  26. Sakaguchi Y, Sekiya I, Yagishita K, Muneta T. Comparison of human stem cells derived from various mesenchymal tissues: Superiority of synovium as a cell source. Arthritis Rheum. 2005;52:2521–2529.
  27. Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002;13:4279–4295.
  28. Fu L, Liu Y, Zhang D, Xie J, Guan H, Shang T. Beneficial effect of human umbilical cord-derived mesenchymal stem cells on an endotoxin-induced rat model of preeclampsia. Exp Ther Med. 2015;10:1851–1856.
  29. Sunil P, Manikandhan R, Muthu M, Abraham S. Stem cell therapy in oral and maxillofacial region: An overview. J Oral Maxillofac Pathol. 2012;16:58–63.