FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Stem Cell Investig / Equine bone marrow-derived mesenchymal stem cells: optimizat...
Stem cell investigation2018; 5; 31; doi: 10.21037/sci.2018.09.01

Equine bone marrow-derived mesenchymal stem cells: optimization of cell density in primary culture.

Abstract: The primary cell seeding density of bone marrow-derived mononuclear cells (BM-MNCs) affects several cellular behaviors, including attachment to the culture dish, proliferation, and differentiation. Methods: The aim of this study was to determine the best density of equine BM-MNCs in primary culture (P0) for obtaining the maximum bone marrow-derived mesenchymal stem cell (BM-MSC) yields at the end of P0. Bone marrow samples of two healthy mares were aspirated. The MNCs were isolated and cultured at different densities (1×10, 2×10, 4×10, 8×10, and 1×10 cells/cm). Within the 7 and 14 days after seeding, the colonies containing more than 15 cells were counted and the percentage of confluency and the number of cells were calculated on day 21. Results: The lowest density of MNCs was associated with the least number of colonies, number of adherent cells, and confluency percentage, whereas the highest density was associated with the maximum number of colonies and confluency percentage (P<0.05). However, the maximum number of cells at the end of P0 was associated with the intermediate (4×10 cells/cm) and the highest concentration (P<0.05). Conclusions: The maximum number of MSCs at the end of P0 was obtained at the densities of 1×10 and, especially, at 4×10 cells/cm.
Get Full Text
Publication Date: 2018-10-09 PubMed ID: 30498742PubMed Central: PMC6232052DOI: 10.21037/sci.2018.09.01Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

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 article presents a study on the optimization of cell density in primary culture for bone marrow-derived stem cells from horses. The study discovered that an intermediate and highest density led to a maximum number of mesenchymal stem cells.

Methodology

  • The researchers aimed at identifying the optimal density for equine bone marrow-derived mononuclear cells (BM-MNCs) in primary culture to achieve maximum mesenchymal stem cell (BM-MSCs) yield at the end of this primary culture period.
  • Bone marrow samples were taken from two healthy mares, and the MNCs within these samples were isolated and cultured at different densities.
  • These densities ranged from 1×10 to 1×10 cells per centimeter squared.
  • Colonies with more than 15 cells were counted 7 and 14 days post-seeding, and the cell numbers and confluency percentage (the percentage of the dish surface covered by cells) were calculated on the 21st day.

Results

  • Results pointed out that the lowest MNCs density led to the least number of colonies and adherent cells, as well as the smallest confluency percentage.
  • Contrarily, the highest MNCs density resulted in the highest number of colonies and confluency percentage.
  • Interestingly, the highest number of cells at the end of the primary culture period (P0) was linked to an intermediate (4×10 cells per centimeter squared) and the highest MNC density.

Conclusion

  • The study concluded that the maximum number of mesenchymal stem cells at the end of the primary culture period was associated with the highest, and particularly, the intermediate cell densities.
  • These findings provide valuable information for researchers working on stem cell research, especially those focusing on equine medicine and regenerative therapy.

Cite This Article

APA
Zahedi M, Parham A, Dehghani H, Kazemi Mehrjerdi H. (2018). Equine bone marrow-derived mesenchymal stem cells: optimization of cell density in primary culture. Stem Cell Investig, 5, 31. https://doi.org/10.21037/sci.2018.09.01

Publication

Stem cell investigation
ISSN: 2306-9759
NlmUniqueID: 101672113
Country: China
Language: English
Volume: 5
Pages: 31
PII: 31

Researcher Affiliations

Zahedi, Morteza
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Parham, Abbas
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
  • Stem Cell Biotechnology and Alterative Regenerative Medicine Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Dehghani, Hesam
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
  • Stem Cell Biotechnology and Alterative Regenerative Medicine Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Kazemi Mehrjerdi, Hossein
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.

Conflict of Interest Statement

Conflicts of Interest: The authors declare that there is no conflict of interest.

References

This article includes 34 references
  1. Ribitsch I, Burk J, Delling U, Geißler C, Gittel C, Jülke H, Brehm W. Basic science and clinical application of stem cells in veterinary medicine.. Adv Biochem Eng Biotechnol 2010;123:219-63.
    pubmed: 20309674doi: 10.1007/10_2010_66google scholar: lookup
  2. 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 Aug;52(8):2521-9.
    doi: 10.1002/art.21212pubmed: 16052568google scholar: lookup
  3. Tondreau T, Meuleman N, Delforge A, Dejeneffe M, Leroy R, Massy M, Mortier C, Bron D, Lagneaux L. Mesenchymal stem cells derived from CD133-positive cells in mobilized peripheral blood and cord blood: proliferation, Oct4 expression, and plasticity.. Stem Cells 2005 Sep;23(8):1105-12.
    doi: 10.1634/stemcells.2004-0330pubmed: 15955825google scholar: lookup
  4. Wintrobe MM, Greer JP. Wintrobe's clinical hematology. Philadelphia: Lippincott Williams & Wilkins; 2009.
  5. Thiriet M. Hematopoiesis. Tissue Functioning and Remodeling in the Circulatory and Ventilatory Systems. Biomathematical and Biomechanical Modeling of the Circulatory and Ventilatory Systems 5: Springer New York; 2013:19-52.
  6. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop Dj, Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.. Cytotherapy 2006;8(4):315-7.
    doi: 10.1080/14653240600855905pubmed: 16923606google scholar: lookup
  7. Lakshmipathy U, Verfaillie C. Stem cell plasticity.. Blood Rev 2005 Jan;19(1):29-38.
    doi: 10.1016/j.blre.2004.03.001pubmed: 15572215google scholar: lookup
  8. Litzke LE, Wagner E, Baumgaertner W, Hetzel U, Josimović-Alasević O, Libera J. Repair of extensive articular cartilage defects in horses by autologous chondrocyte transplantation.. Ann Biomed Eng 2004 Jan;32(1):57-69.
    doi: 10.1023/B:ABME.0000007791.81433.1apubmed: 14964722google scholar: lookup
  9. Pacini S, Spinabella S, Trombi L, Fazzi R, Galimberti S, Dini F, Carlucci F, Petrini M. Suspension of bone marrow-derived undifferentiated mesenchymal stromal cells for repair of superficial digital flexor tendon in race horses.. Tissue Eng 2007 Dec;13(12):2949-55.
    doi: 10.1089/ten.2007.0108pubmed: 17919069google scholar: lookup
  10. Smith RK. Mesenchymal stem cell therapy for equine tendinopathy.. Disabil Rehabil 2008;30(20-22):1752-8.
    doi: 10.1080/09638280701788241pubmed: 18608378google scholar: lookup
  11. Augello A, Tasso R, Negrini SM, Cancedda R, Pennesi G. Cell therapy using allogeneic bone marrow mesenchymal stem cells prevents tissue damage in collagen-induced arthritis.. Arthritis Rheum 2007 Apr;56(4):1175-86.
    doi: 10.1002/art.22511pubmed: 17393437google scholar: lookup
  12. Adel N, Gabr H. Stem cell therapy of acute spinal cord injury in dogs. Third World Congress of Renerative Medicine Regen Med 2007: Future Medicine.
  13. Chopp M, Zhang XH, Li Y, Wang L, Chen J, Lu D, Lu M, Rosenblum M. Spinal cord injury in rat: treatment with bone marrow stromal cell transplantation.. Neuroreport 2000 Sep 11;11(13):3001-5.
    doi: 10.1097/00001756-200009110-00035pubmed: 11006983google scholar: lookup
  14. Kallis YN, Alison MR, Forbes SJ. Bone marrow stem cells and liver disease.. Gut 2007 May;56(5):716-24.
    doi: 10.1136/gut.2006.098442pmc: PMC1942133pubmed: 17145739google scholar: lookup
  15. Kraus KH, Kirker-Head C. Mesenchymal stem cells and bone regeneration.. Vet Surg 2006 Apr;35(3):232-42.
    doi: 10.1111/j.1532-950X.2006.00142.xpubmed: 16635002google scholar: lookup
  16. Borjesson DL, Peroni JF. The regenerative medicine laboratory: facilitating stem cell therapy for equine disease.. Clin Lab Med 2011 Mar;31(1):109-23.
    doi: 10.1016/j.cll.2010.12.001pubmed: 21295725google scholar: lookup
  17. Michler RE. Stem cell therapy for heart failure.. Methodist Debakey Cardiovasc J 2013 Oct-Dec;9(4):187-94.
    doi: 10.14797/mdcj-9-4-187pmc: PMC3846071pubmed: 24298308google scholar: lookup
  18. Friedenstein AJ, Chailakhjan RK, Lalykina KS. The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells.. Cell Tissue Kinet 1970 Oct;3(4):393-403.
    doi: 10.1111/j.1365-2184.1970.tb00347.xpubmed: 5523063google scholar: lookup
  19. Vidal MA, Kilroy GE, Johnson JR, Lopez MJ, Moore RM, Gimble JM. Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity.. Vet Surg 2006 Oct;35(7):601-10.
    doi: 10.1111/j.1532-950X.2006.00197.xpubmed: 17026544google scholar: lookup
  20. Kulterer B, Friedl G, Jandrositz A, Sanchez-Cabo F, Prokesch A, Paar C, Scheideler M, Windhager R, Preisegger KH, Trajanoski Z. Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation.. BMC Genomics 2007 Mar 12;8:70.
    doi: 10.1186/1471-2164-8-70pmc: PMC1829400pubmed: 17352823google scholar: lookup
  21. Martin DR, Cox NR, Hathcock TL, Niemeyer GP, Baker HJ. Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow.. Exp Hematol 2002 Aug;30(8):879-86.
    doi: 10.1016/S0301-472X(02)00864-0pubmed: 12160839google scholar: lookup
  22. Kadiyala S, Young RG, Thiede MA, Bruder SP. Culture expanded canine mesenchymal stem cells possess osteochondrogenic potential in vivo and in vitro.. Cell Transplant 1997 Mar-Apr;6(2):125-34.
    doi: 10.1177/096368979700600206pubmed: 9142444google scholar: lookup
  23. 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.
    doi: 10.1126/science.284.5411.143pubmed: 10102814google scholar: lookup
  24. Bourzac C, Smith LC, Vincent P, Beauchamp G, Lavoie JP, Laverty S. Isolation of equine bone marrow-derived mesenchymal stem cells: a comparison between three protocols.. Equine Vet J 2010 Sep;42(6):519-27.
    doi: 10.1111/j.2042-3306.2010.00098.xpubmed: 20716192google scholar: lookup
  25. Stanley AJ, Banks RE, Southgate J, Selby PJ. Effect of cell density on the expression of adhesion molecules and modulation by cytokines.. Cytometry 1995 Dec 1;21(4):338-43.
    doi: 10.1002/cyto.990210405pubmed: 8608731google scholar: lookup
  26. Ben-Ze'ev A, Robinson GS, Bucher NL, Farmer SR. Cell-cell and cell-matrix interactions differentially regulate the expression of hepatic and cytoskeletal genes in primary cultures of rat hepatocytes.. Proc Natl Acad Sci U S A 1988 Apr;85(7):2161-5.
    doi: 10.1073/pnas.85.7.2161pmc: PMC279949pubmed: 3353374google scholar: lookup
  27. Albelda SM, Buck CA. Integrins and other cell adhesion molecules.. FASEB J 1990 Aug;4(11):2868-80.
    doi: 10.1096/fasebj.4.11.2199285pubmed: 2199285google scholar: lookup
  28. De Schauwer C, Meyer E, Van de Walle GR, Van Soom A. Markers of stemness in equine mesenchymal stem cells: a plea for uniformity.. Theriogenology 2011 May;75(8):1431-43.
    doi: 10.1016/j.theriogenology.2010.11.008pubmed: 21196039google scholar: lookup
  29. Piedimonte G, Borghetti AF, Guidotti GG. Effect of cell density on growth rate and amino acid transport in simian virus 40-transformed 3T3 cells.. Cancer Res 1982 Nov;42(11):4690-3.
    pubmed: 6290044
  30. Pardee AB. The cell surface and fibroblast proliferation some current research trends.. Biochim Biophys Acta 1975 Jul 11;417(2):153-72.
    pubmed: 167866doi: 10.1016/0304-419x(75)90003-7google scholar: lookup
  31. Alipour F, Parham A, Kazemi Mehrjerdi H, Dehghani H. Equine adipose-derived mesenchymal stem cells: phenotype and growth characteristics, gene expression profile and differentiation potentials.. Cell J 2015 Winter;16(4):456-65.
    pmc: PMC4297484pubmed: 25685736doi: 10.22074/cellj.2015.491google scholar: lookup
  32. Giovannini S, Brehm W, Mainil-Varlet P, Nesic D. Multilineage differentiation potential of equine blood-derived fibroblast-like cells.. Differentiation 2008 Feb;76(2):118-29.
    doi: 10.1111/j.1432-0436.2007.00207.xpubmed: 17697129google scholar: lookup
  33. Ibrahim S, Steinbach F. Non-HLDA8 animal homologue section anti-leukocyte mAbs tested for reactivity with equine leukocytes.. Vet Immunol Immunopathol 2007 Sep 15;119(1-2):81-91.
    doi: 10.1016/j.vetimm.2007.06.033pubmed: 17692930google scholar: lookup
  34. Radcliffe CH, Flaminio MJ, Fortier LA. Temporal analysis of equine bone marrow aspirate during establishment of putative mesenchymal progenitor cell populations.. Stem Cells Dev 2010 Feb;19(2):269-82.
    doi: 10.1089/scd.2009.0091pmc: PMC3138180pubmed: 19604071google scholar: lookup

Citations

This article has been cited 4 times.
  1. Che L, Zhu C, Huang L, Xu H, Ma X, Luo X, He H, Zhang T, Wang N. Ginsenoside Rg2 Promotes the Proliferation and Stemness Maintenance of Porcine Mesenchymal Stem Cells through Autophagy Induction.. Foods 2023 Mar 2;12(5).
    doi: 10.3390/foods12051075pubmed: 36900592google scholar: lookup
  2. Nagai H, Miwa A, Yoneda K, Fujisawa K, Takami T. Optimizing the Seeding Density of Human Mononuclear Cells to Improve the Purity of Highly Proliferative Mesenchymal Stem Cells.. Bioengineering (Basel) 2023 Jan 11;10(1).
    doi: 10.3390/bioengineering10010102pubmed: 36671674google scholar: lookup
  3. Smieszek A, Marcinkowska K, Pielok A, Sikora M, Valihrach L, Carnevale E, Marycz K. Obesity Affects the Proliferative Potential of Equine Endometrial Progenitor Cells and Modulates Their Molecular Phenotype Associated with Mitochondrial Metabolism.. Cells 2022 Apr 24;11(9).
    doi: 10.3390/cells11091437pubmed: 35563743google scholar: lookup
  4. Bagge J, MacLeod JN, Berg LC. Cellular Proliferation of Equine Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem Cells Decline With Increasing Donor Age.. Front Vet Sci 2020;7:602403.
    doi: 10.3389/fvets.2020.602403pubmed: 33363241google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.