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Home / Equine Research Bank / Vet Res Commun / Characterization of adipose-derived equine and canine mesenc...
Veterinary research communications2011; 35(8); 487-499; doi: 10.1007/s11259-011-9492-8

Characterization of adipose-derived equine and canine mesenchymal stem cells after incubation in agarose-hydrogel.

Abstract: Adult stem cells are of particular interest for the therapeutic approach in the field of regenerative medicine. Due to their ease of harvest, adipose-derived mesenchymal stem cells (ASCs) are an attractive stem cell source that has become increasingly popular. Critical aspects of applied cell therapies are the circumstances of transport from the laboratory towards the site of operation and cell delivery into the desired area. With regard to these issues, agarose-hydrogel was analyzed as a cell carrier matrix of equine and canine ASCs in vitro, which can be used for minimally invasive application. Isolated ASCs were expanded and 2.5 × 10(6) cells were combined with agarose-hydrogel to build a 0.4% hydrogel-cell solution which was stored at two temperatures (room temperature (RT) vs. 37 °C). Cell viability was investigated (live-dead assay) at different time points (0, 1, 6 and 24 h) in order to determine i) the effect of different temperatures on the cell survival as well as ii) the maximum possible time span before implantation. CFU-assay and WST-1 assay were performed after 24 h incubation in agarose-hydrogel and the cells were induced into adipogenic and osteogenic differentiation to analyze the effects of the incubation on the cell behaviour. No negative effect of the agarose-hydrogel incubation was determined on the different species' cell behaviour at either RT or 37 °C with any of the assays used. We can recommend agarose-hydrogel as a cell carrier for cell implantation with a storage period of up to 24 h at room temperature or at 37 °C prior to implantation.
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Publication Date: 2011-07-15 PubMed ID: 21755422DOI: 10.1007/s11259-011-9492-8Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates the viability of adipose-derived (fat-tissue based) equine and canine stem cells when stored in an agarose-hydrogel solution before cell-therapy based medical treatment. The main findings suggest that these stem cells can be safely stored in this solution for up to 24 hours without negatively affecting their behavior.

Overview of the Study

  • This study was conducted to assess the efficiency of agarose-hydrogel as a cell carrier for adipose-derived mesenchymal stem cells (ASCs). These types of stem cells are often used in regenerative medicine due to their easy harvesting procedure.
  • The researchers focused on the delivery and transportation of these cells from the lab to the operation site, which can often affect their viability and effectiveness.
  • Agarose-hydrogel was selected as a medium to carry these cells as it enables minimally invasive application.

Methodology and Execution

  • ASCs were isolated, expanded and combined with an agarose-hydrogel to create a 0.4% hydrogel-cell solution.
  • This solution was then stored at two different temperatures, room temperature (RT) and 37 °C, to measure the impact of temperature on cell survival.
  • A live-dead assay was used to determine cell viability at different time points (0, 1, 6 and 24 hours). This was done to understand the maximum length of time cells can be stored before implantation.
  • Other assays like the CFU-assay and WST-1 assay were also performed after a 24-hour incubation period in the agarose-hydrogel to study its effects on cell behaviour. Additionally, the cells were induced to differentiate into fat and bone cells to further study the impact of the agarose-hydrogel incubation.

Key Findings and Implications

  • From the different assays and tests conducted, the researchers found that the agarose-hydrogel incubation had no adverse effects on either equine or canine stem cells at both tested temperatures.
  • Therefore, the study concludes that agarose-hydrogel could be an effective cell carrier medium for stem cells with a safe storage period of up to 24 hours at both room temperature and 37 °C before implantation.

Cite This Article

APA
Schwarz C, Leicht U, Drosse I, Ulrich V, Luibl V, Schieker M, Röcken M. (2011). Characterization of adipose-derived equine and canine mesenchymal stem cells after incubation in agarose-hydrogel. Vet Res Commun, 35(8), 487-499. https://doi.org/10.1007/s11259-011-9492-8

Publication

Veterinary research communications
ISSN: 1573-7446
NlmUniqueID: 8100520
Country: Switzerland
Language: English
Volume: 35
Issue: 8
Pages: 487-499

Researcher Affiliations

Schwarz, Christina
  • Laboratory for Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig Maximilians University, Nussbaumstr. 20, 80336, Munich, Germany.
Leicht, Uta
    Drosse, Inga
      Ulrich, Veronika
        Luibl, Volker
          Schieker, Matthias
            Röcken, Michael

              MeSH Terms

              • Adipose Tissue / cytology
              • Animals
              • Cell Culture Techniques
              • Cell Differentiation
              • Dogs
              • Female
              • Horses
              • Hydrogels / chemistry
              • Male
              • Mesenchymal Stem Cells / cytology
              • Mesenchymal Stem Cells / physiology
              • Sepharose / chemistry

              References

              This article includes 38 references
              1. Schieker M, Mutschler W. [Bridging posttraumatic bony defects. Established and new methods].. Unfallchirurg 2006 Sep;109(9):715-32.
                pubmed: 16941096doi: 10.1007/s00113-006-1152-zgoogle scholar: lookup
              2. Giannoudis PV, Einhorn TA, Marsh D. Fracture healing: the diamond concept.. Injury 2007 Sep;38 Suppl 4:S3-6.
                pubmed: 18224731doi: 10.1016/s0020-1383(08)70003-2google scholar: lookup
              3. Rimondini L, Mele S. Stem cell technologies for tissue regeneration in dentistry.. Minerva Stomatol 2009 Oct;58(10):483-500.
                pubmed: 19893474
              4. Gimble JM. Adipose tissue-derived therapeutics.. Expert Opin Biol Ther 2003 Aug;3(5):705-13.
                pubmed: 12880371doi: 10.1517/14712598.3.5.705google scholar: lookup
              5. Awad HA, Butler DL, Boivin GP, Smith FN, Malaviya P, Huibregtse B, Caplan AI. Autologous mesenchymal stem cell-mediated repair of tendon.. Tissue Eng 1999 Jun;5(3):267-77.
                pubmed: 10434073doi: 10.1089/ten.1999.5.267google scholar: lookup
              6. Agashi K, Chau DY, Shakesheff KM. The effect of delivery via narrow-bore needles on mesenchymal cells.. Regen Med 2009 Jan;4(1):49-64.
                pubmed: 19105616doi: 10.2217/17460751.4.1.49google scholar: lookup
              7. Caplan AI. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine.. J Cell Physiol 2007 Nov;213(2):341-7.
                pubmed: 17620285doi: 10.1002/jcp.21200google scholar: lookup
              8. Awad HA, Boivin GP, Dressler MR, Smith FN, Young RG, Butler DL. Repair of patellar tendon injuries using a cell-collagen composite.. J Orthop Res 2003 May;21(3):420-31.
                pubmed: 12706014doi: 10.1016/S0736-0266(02)00163-8google scholar: lookup
              9. Lacitignola L, Crovace A, Rossi G, Francioso E. Cell therapy for tendinitis, experimental and clinical report.. Vet Res Commun 2008 Sep;32 Suppl 1:S33-8.
                pubmed: 18686004doi: 10.1007/s11259-008-9085-3google scholar: lookup
              10. Morrison SJ, Shah NM, Anderson DJ. Regulatory mechanisms in stem cell biology.. Cell 1997 Feb 7;88(3):287-98.
                pubmed: 9039255doi: 10.1016/s0092-8674(00)81867-xgoogle scholar: lookup
              11. Schieker M, Seitz S, Gülkan H, Nentwich M, Horvath G, Regauer M, Milz S, Mutschler W. [Tissue engineering of bone. Integration and migration of human mesenchymal stem cells in colonized contructs in a murine model].. Orthopade 2004 Dec;33(12):1354-60.
                pubmed: 15551050doi: 10.1007/s00132-004-0740-2google scholar: lookup
              12. Duggal S, Brinchmann JE. Importance of serum source for the in vitro replicative senescence of human bone marrow derived mesenchymal stem cells.. J Cell Physiol 2011 Nov;226(11):2908-15.
                pubmed: 21302288doi: 10.1002/jcp.22637google scholar: lookup
              13. Schnabel LV, Lynch ME, van der Meulen MC, Yeager AE, Kornatowski MA, Nixon AJ. Mesenchymal stem cells and insulin-like growth factor-I gene-enhanced mesenchymal stem cells improve structural aspects of healing in equine flexor digitorum superficialis tendons.. J Orthop Res 2009 Oct;27(10):1392-8.
                pubmed: 19350658doi: 10.1002/jor.20887google scholar: lookup
              14. Drosse I, Volkmer E, Capanna R, De Biase P, Mutschler W, Schieker M. Tissue engineering for bone defect healing: an update on a multi-component approach.. Injury 2008 Sep;39 Suppl 2:S9-20.
                pubmed: 18804579doi: 10.1016/S0020-1383(08)70011-1google scholar: lookup
              15. Anderson GB, Foote RH. Development of rabbit embryos in vitro and in vivo following storage of the two-cell stage at 10 degrees C.. J Reprod Fertil 1975 Oct;45(1):151-3.
                pubmed: 1238563doi: 10.1530/jrf.0.0450151google scholar: lookup
              16. Park JH, Chung BG, Lee WG, Kim J, Brigham MD, Shim J, Lee S, Hwang CM, Durmus NG, Demirci U, Khademhosseini A. Microporous cell-laden hydrogels for engineered tissue constructs.. Biotechnol Bioeng 2010 May 1;106(1):138-48.
                pubmed: 20091766doi: 10.1002/bit.22667google scholar: lookup
              17. Davatchi F, Abdollahi BS, Mohyeddin M, Shahram F, Nikbin B. Mesenchymal stem cell therapy for knee osteoarthritis. Preliminary report of four patients.. Int J Rheum Dis 2011 May;14(2):211-5.
                pubmed: 21518322doi: 10.1111/j.1756-185X.2011.01599.xgoogle scholar: lookup
              18. Dzik WH. The air we breathe: three vital respiratory gases and the red blood cell: oxygen, nitric oxide, and carbon dioxide.. Transfusion 2011 Apr;51(4):676-85.
                pubmed: 21496039doi: 10.1111/j.1537-2995.2011.03114.xgoogle scholar: lookup
              19. Zuidema JM, Pap MM, Jaroch DB, Morrison FA, Gilbert RJ. Fabrication and characterization of tunable polysaccharide hydrogel blends for neural repair.. Acta Biomater 2011 Apr;7(4):1634-43.
                pubmed: 21130187doi: 10.1016/j.actbio.2010.11.039google scholar: lookup
              20. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies.. Tissue Eng 2001 Apr;7(2):211-28.
                pubmed: 11304456doi: 10.1089/107632701300062859google scholar: lookup
              21. Del Bue M, Riccò S, Ramoni R, Conti V, Gnudi G, Grolli S. Equine adipose-tissue derived mesenchymal stem cells and platelet concentrates: their association in vitro and in vivo.. Vet Res Commun 2008 Sep;32 Suppl 1:S51-5.
                pubmed: 18683070doi: 10.1007/s11259-008-9093-3google scholar: lookup
              22. Juncosa-Melvin N, Boivin GP, Gooch C, Galloway MT, West JR, Dunn MG, Butler DL. The effect of autologous mesenchymal stem cells on the biomechanics and histology of gel-collagen sponge constructs used for rabbit patellar tendon repair.. Tissue Eng 2006 Feb;12(2):369-79.
                pubmed: 16548695doi: 10.1089/ten.2006.12.369google scholar: lookup
              23. Yamamoto N, Akamatsu H, Hasegawa S, Yamada T, Nakata S, Ohkuma M, Miyachi E, Marunouchi T, Matsunaga K. Isolation of multipotent stem cells from mouse adipose tissue.. J Dermatol Sci 2007 Oct;48(1):43-52.
                pubmed: 17644316doi: 10.1016/j.jdermsci.2007.05.015google scholar: lookup
              24. Meuleman N, Tondreau T, Delforge A, Dejeneffe M, Massy M, Libertalis M, Bron D, Lagneaux L. Human marrow mesenchymal stem cell culture: serum-free medium allows better expansion than classical alpha-MEM medium.. Eur J Haematol 2006 Apr;76(4):309-16.
                pubmed: 16519702doi: 10.1111/j.1600-0609.2005.00611.xgoogle scholar: lookup
              25. Tapp H, Hanley EN Jr, Patt JC, Gruber HE. Adipose-derived stem cells: characterization and current application in orthopaedic tissue repair.. Exp Biol Med (Maywood) 2009 Jan;234(1):1-9.
                pubmed: 19109553doi: 10.3181/0805/MR-170google scholar: lookup
              26. Volkmer E, Drosse I, Otto S, Stangelmayer A, Stengele M, Kallukalam BC, Mutschler W, Schieker M. Hypoxia in static and dynamic 3D culture systems for tissue engineering of bone.. Tissue Eng Part A 2008 Aug;14(8):1331-40.
                pubmed: 18601588doi: 10.1089/ten.tea.2007.0231google scholar: lookup
              27. 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 Apr 2;284(5411):143-7.
                pubmed: 10102814doi: 10.1126/science.284.5411.143google scholar: lookup
              28. Spitkovsky D, Hescheler J. Adult mesenchymal stromal stem cells for therapeutic applications.. Minim Invasive Ther Allied Technol 2008;17(2):79-90.
                pubmed: 18465443doi: 10.1080/13645700801969758google scholar: lookup
              29. Fedorovich NE, Alblas J, de Wijn JR, Hennink WE, Verbout AJ, Dhert WJ. Hydrogels as extracellular matrices for skeletal tissue engineering: state-of-the-art and novel application in organ printing.. Tissue Eng 2007 Aug;13(8):1905-25.
                pubmed: 17518748doi: 10.1089/ten.2006.0175google scholar: lookup
              30. Guilak F, Lott KE, Awad HA, Cao Q, Hicok KC, Fermor B, Gimble JM. Clonal analysis of the differentiation potential of human adipose-derived adult stem cells.. J Cell Physiol 2006 Jan;206(1):229-37.
                pubmed: 16021633doi: 10.1002/jcp.20463google scholar: lookup
              31. Shahdadfar A, Frønsdal K, Haug T, Reinholt FP, Brinchmann JE. In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability.. Stem Cells 2005 Oct;23(9):1357-66.
                pubmed: 16081661doi: 10.1634/stemcells.2005-0094google scholar: lookup
              32. Watanabe J, Kashii M, Hirao M, Oka K, Sugamoto K, Yoshikawa H, Akashi M. Quick-forming hydroxyapatite/agarose gel composites induce bone regeneration.. J Biomed Mater Res A 2007 Dec 1;83(3):845-52.
                pubmed: 17559128doi: 10.1002/jbm.a.31435google scholar: lookup
              33. Chase LG, Lakshmipathy U, Solchaga LA, Rao MS, Vemuri MC. A novel serum-free medium for the expansion of human mesenchymal stem cells.. Stem Cell Res Ther 2010 Apr 2;1(1):8.
                pubmed: 20504289doi: 10.1186/scrt8google scholar: lookup
              34. Goh JC, Ouyang HW, Teoh SH, Chan CK, Lee EH. Tissue-engineering approach to the repair and regeneration of tendons and ligaments.. Tissue Eng 2003;9 Suppl 1:S31-44.
                pubmed: 14511469doi: 10.1089/10763270360696969google scholar: lookup
              35. Nicodemus GD, Bryant SJ. Cell encapsulation in biodegradable hydrogels for tissue engineering applications.. Tissue Eng Part B Rev 2008 Jun;14(2):149-65.
                pubmed: 18498217doi: 10.1089/ten.teb.2007.0332google scholar: lookup
              36. Sakai S, Hashimoto I, Kawakami K. Synthesis of an agarose-gelatin conjugate for use as a tissue engineering scaffold.. J Biosci Bioeng 2007 Jan;103(1):22-6.
                pubmed: 17298896doi: 10.1263/jbb.103.22google scholar: lookup
              37. Schumacher A, Fischer B. Influence of visible light and room temperature on cell proliferation in preimplantation rabbit embryos.. J Reprod Fertil 1988 Sep;84(1):197-204.
                pubmed: 3184041doi: 10.1530/jrf.0.0840197google scholar: lookup
              38. Anderson GB, Foote RH. Effects of low temperature upon subsequent nucleic acid and protein synthesis of rabbit embryos.. Exp Cell Res 1975 Jan;90(1):73-8.
                pubmed: 1122946doi: 10.1016/0014-4827(75)90358-4google scholar: lookup

              Citations

              This article has been cited 7 times.
              1. Kemilew J, Sobczyńska-Rak A, Żylińska B, Szponder T, Nowicka B, Urban B. The Use of Allogenic Stromal Vascular Fraction (SVF) Cells in Degenerative Joint Disease of the Spine in Dogs. In Vivo 2019 Jul-Aug;33(4):1109-1117.
                doi: 10.21873/invivo.11580pubmed: 31280199google scholar: lookup
              2. Zanna N, Focaroli S, Merlettini A, Gentilucci L, Teti G, Falconi M, Tomasini C. Thixotropic Peptide-Based Physical Hydrogels Applied to Three-Dimensional Cell Culture. ACS Omega 2017 May 31;2(5):2374-2381.
                doi: 10.1021/acsomega.7b00322pubmed: 30023662google scholar: lookup
              3. Tidd N, Michelsen J, Hilbert B, Quinn JC. Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells. Int J Mol Sci 2017 Apr 12;18(4).
                doi: 10.3390/ijms18040819pubmed: 28417917google scholar: lookup
              4. Xu G, Yin F, Wu H, Hu X, Zheng L, Zhao J. In vitro ovarian cancer model based on three-dimensional agarose hydrogel. J Tissue Eng 2014;5:2041731413520438.
                doi: 10.1177/2041731413520438pubmed: 24551446google scholar: lookup
              5. Astor DE, Hoelzler MG, Harman R, Bastian RP. Patient factors influencing the concentration of stromal vascular fraction (SVF) for adipose-derived stromal cell (ASC) therapy in dogs. Can J Vet Res 2013 Jul;77(3):177-82.
                pubmed: 24101793
              6. Volkmer E, Leicht U, Moritz M, Schwarz C, Wiese H, Milz S, Matthias P, Schloegl W, Friess W, Goettlinger M, Augat P, Schieker M. Poloxamer-based hydrogels hardening at body core temperature as carriers for cell based therapies: in vitro and in vivo analysis. J Mater Sci Mater Med 2013 Sep;24(9):2223-34.
                doi: 10.1007/s10856-013-4966-6pubmed: 23712537google scholar: lookup
              7. Oh HJ, Park EJ, Lee SY, Soh JW, Kong IS, Choi SW, Ra JC, Kang SK, Lee BC. Comparison of cell proliferation and epigenetic modification of gene expression patterns in canine foetal fibroblasts and adipose tissue-derived mesenchymal stem cells. Cell Prolif 2012 Oct;45(5):438-44.
                doi: 10.1111/j.1365-2184.2012.00838.xpubmed: 22925503google scholar: lookup
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