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Isolation and culture of primary rat adipose derived stem cells using porous biopolymer microcarriers

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Adipose-derived stem cells (ADSCs) are an attractive source of material for mesenchymal stem cell research due to the abundance of adipose and relative ease of access compared with bone marrow. A key consideration for research is whether cell isolation methods can be improved, to reduce the process steps needed to isolate and expand cell material. In the current study, we used macroporous biopolymer microcarriers to isolate primary ADSCs. We found that the method was capable of isolating ADSCs that were subsequently capable of being transferred to culture dishes and expanded in vitro. Moreover, flow cytometry revealed that they expressed typical stem cell markers and were capable of undergoing tri-lineage differentiation. In summary, it is feasible to use biopolymer microcarriers for retrieval of viable ADSCs that retain identity markers of stem cell function.

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References

  1. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001;7:211–228.

    Article  CAS  PubMed  Google Scholar 

  2. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002;13:4279–4295.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Hong L, Colpan A, Peptan IA, Daw J, George A, Evans CA. 17-Beta estradiol enhances osteogenic and adipogenic differentiation of human adipose-derived stromal cells. Tissue Eng 2007;13:1197–1203.

    Article  CAS  PubMed  Google Scholar 

  4. Minteer DM, Young MT, Lin YC, Over PJ, Rubin JP, Gerlach JC, et al. Analysis of type II diabetes mellitus adipose-derived stem cells for tissue engineering applications. J Tissue Eng 2015;6:2041731415579215.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Davies OG, Cooper PR, Shelton RM, Smith AJ, Scheven BA. Isolation of adipose and bone marrow mesenchymal stem cells using CD29 and CD90 modifies their capacity for osteogenic and adipogenic differentiation. J Tissue Eng 2015;6:2041731415592356.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Anghileri E, Marconi S, Pignatelli A, Cifelli P, Galié M, Sbarbati A, et al. Neuronal differentiation potential of human adipose-derived mesenchymal stem cells. Stem Cells Dev 2008;17:909–916.

    Article  CAS  PubMed  Google Scholar 

  7. Dugan JM, Cartmell SH, Gough JE. Uniaxial cyclic strain of human adipose-derived mesenchymal stem cells and C2C12 myoblasts in coculture. J Tissue Eng 2014;5:2041731414530138.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Araña M, Gavira JJ, Peña E, González A, Abizanda G, Cilla M, et al. Epicardial delivery of collagen patches with adipose-derived stem cells in rat and minipig models of chronic myocardial infarction. Biomaterials 2014; 35:143–151.

    Article  PubMed  Google Scholar 

  9. Liang CZ, Li H, Tao YQ, Peng LH, Gao JQ, Wu JJ, et al. Dual release of dexamethasone and TGF-ß3 from polymeric microspheres for stem cell matrix accumulation in a rat disc degeneration model. Acta Biomater 2013;9:9423–9433.

    Article  CAS  PubMed  Google Scholar 

  10. Zhou Y, Yan Z, Zhang H, Lu W, Liu S, Huang X, et al. Expansion and delivery of adipose-derived mesenchymal stem cells on three microcarriers for soft tissue regeneration. Tissue Eng Part A 2011;17:2981–2997.

    Article  CAS  PubMed  Google Scholar 

  11. Zhao Y, Lin H, Zhang J, Chen B, Sun W, Wang X, et al. Crosslinked threedimensional demineralized bone matrix for the adipose-derived stromal cell proliferation and differentiation. Tissue Eng Part A 2009; 15:13–21.

    Article  CAS  PubMed  Google Scholar 

  12. Aguena M, Fanganiello RD, Tissiani LA, Ishiy FA, Atique R, Alonso N, et al. Optimization of parameters for a more efficient use of adipose-derived stem cells in regenerative medicine therapies. Stem Cells Int 2012; 2012:303610.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Ahmadi N, Razavi S, Kazemi M, Oryan S. Stability of neural differentia tion in human adipose derived stem cells by two induction protocols. Tissue Cell 2012;44:87–94.

    Article  CAS  PubMed  Google Scholar 

  14. Ghorbani A, Jalali SA, Varedi M. Isolation of adipose tissue mesenchymal stem cells without tissue destruction: a non-enzymatic method. Tissue Cell 2014;46:54–58.

    Article  CAS  PubMed  Google Scholar 

  15. Guedes JC, Park JH, Lakhkar NJ, Kim HW, Knowles JC, Wall IB. TiO2-doped phosphate glass microcarriers: a stable bioactive substrate for expansion of adherent mammalian cells. J Biomater Appl 2013;28:3–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Park JH, Pérez RA, Jin GZ, Choi SJ, Kim HW, Wall IB. Microcarriers designed for cell culture and tissue engineering of bone. Tissue Eng Part B Rev 2013;19:172–190.

    Article  CAS  PubMed  Google Scholar 

  17. Lakhkar NJ, M Day R, Kim HW, Ludka K, Mordan NJ, Salih V, et al. Titanium phosphate glass microcarriers induce enhanced osteogenic cell proliferation and human mesenchymal stem cell protein expression. J Tissue Eng 2015;6:2041731415617741.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Jin GZ, Park JH, Lee EJ, Wall IB, Kim HW. Utilizing PCL microcarriers for high-purity isolation of primary endothelial cells for tissue engineering. Tissue Eng Part C Methods 2014;20:761–768.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Nagle RB. Intermediate filaments: a review of the basic biology. Am J Surg Pathol 1988;12 Suppl 1:4–16.

    PubMed  Google Scholar 

  20. de Macedo Braga LM, Lacchini S, Schaan BD, Rodrigues B, Rosa K, De Angelis K, et al. In situ delivery of bone marrow cells and mesenchymal stem cells improves cardiovascular function in hypertensive rats submitted to myocardial infarction. J Biomed Sci 2008;15:365–374.

    Article  PubMed  Google Scholar 

  21. Hong SJ, Yu HS, Kim HW. Tissue engineering polymeric microcarriers with macroporous morphology and bone-bioactive surface. Macromol Biosci 2009;9:639–645.

    Article  CAS  PubMed  Google Scholar 

  22. Lee HH, Hong SJ, Kim CH, Kim EC, Jang JH, Shin HI, et al. Preparation of hydroxyapatite spheres with an internal cavity as a scaffold for hard tissue regeneration. J Mater Sci Mater Med 2008;19:3029–3034.

    Article  CAS  PubMed  Google Scholar 

  23. Park JH, Lee EJ, Knowles JC, Kim HW. Preparation of in situ hardening composite microcarriers: calcium phosphate cement combined with alginate for bone regeneration. J Biomater Appl 2014;28:1079–1084.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Reckhenrich AK, Kirsch BM, Wahl EA, Schenck TL, Rezaeian F, Harder Y, et al. Surgical sutures filled with adipose-derived stem cells promote wound healing. PLoS One 2014;9:e91169.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Guyette JP, Fakharzadeh M, Burford EJ, Tao ZW, Pins GD, Rolle MW, et al. A novel suture-based method for efficient transplantation of stem cells. J Biomed Mater Res A 2013;101:809–818.

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Institute of Tissue Regeneration Engineering (ITREN), Dankook University, 119 Dandae-ro, Dongnam-gu, Cheonan, 31116, Korea

    Guang-Zhen Jin, Jeong-Hui Park, Ivan Wall & Hae-Won Kim

  2. Department of Nanobiomedical Science & BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan, Korea

    Guang-Zhen Jin, Jeong-Hui Park & Hae-Won Kim

  3. Department of Biochemical Engineering, University College London, Gordon Street, London, WC1H 0AH, UK

    Ivan Wall

  4. Department of Biomaterials Science, Dankook University Dental College, Cheonan, Korea

    Hae-Won Kim

Authors
  1. Guang-Zhen Jin
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  2. Jeong-Hui Park
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  3. Ivan Wall
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  4. Hae-Won Kim
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Correspondence to Ivan Wall or Hae-Won Kim.

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Jin, GZ., Park, JH., Wall, I. et al. Isolation and culture of primary rat adipose derived stem cells using porous biopolymer microcarriers. Tissue Eng Regen Med 13, 242–250 (2016). https://doi.org/10.1007/s13770-016-0040-z

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  • DOI: https://doi.org/10.1007/s13770-016-0040-z

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