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BMC veterinary research2018; 14(1); 247; doi: 10.1186/s12917-018-1572-3

Growth and function of equine endothelial colony forming cells labeled with semiconductor quantum dots.

Abstract: Endothelial progenitor cells (EPCs) contribute to neovascularization and vascular repair in vivo and are attractive for clinical use in ischemic disease. Tracking of stem and progenitor cells is essential to determine engraftment after administration. Semiconductor quantum dots (QD) are promising for cell labeling due to their ease of uptake by many cell lines and their continued presence after many cell generations. The purpose of this study was to evaluate function and growth of equine EPCs after QD labeling. Additionally, this study evaluated the duration of QD label retention and mechanisms of QD label loss. Results: Endothelial colony forming cells (ECFCs) from adult horses (N = 3) were employed for this study, with QD labeled and unlabeled ECFCs tested from each horse. Cell proliferation of ECFCs labeled with QD at 20 nM was quantified by comparing the number of cell doublings per day (NCD) and the population doubling time (PDT) in labeled and unlabeled cells. Function of labeled and unlabeled ECFCs was assessed by comparing uptake of acetylated low-density lipoprotein (DiO-Ac-LDL) and tubule formation on growth factor containing matrix. Cell proliferation was not impacted by QD labeling; both NCD (p = 0. 95) and PDT (P = 0. 91) did not differ between unlabeled and QD labeled cells. Function of ECFCs assessed by DiO-Ac-LDL and tubule formation was also not different between unlabeled and QD labeled cells (P = 0. 33 and P = 0. 52, respectively). ECFCs retained their QD labeling over 7 passages with both 5 nM and 20 nM label concentrations. Reduction in label intensity was observed over time, and the mechanism was determined to be cell division. Conclusions: Equine ECFCs are effectively labeled with QD, and QD concentrations up to 20 nM do not affect cell growth or function. QD label loss is a result of cell division. The use of QD labeling with equine EPCs may be an ideal way to track engraftment of EPCs for in vivo applications.
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Publication Date: 2018-08-23 PubMed ID: 30139355PubMed Central: PMC6107939DOI: 10.1186/s12917-018-1572-3Google Scholar: Lookup
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Summary

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The research article investigates the effect of semiconductor quantum dots (QD) labeling on the growth and function of equine Endothelial Colony Forming Cells (ECFCs). The study found that quantum dots don’t affect cell growth or function, with cell division identified as the reason for quantum dots label loss. The researchers suggest that quantum dots could be an effective tool for tracing ECFCs in clinical applications.

Objective

The main goal of this research is to assess the impact of Quantum Dots (QD) labeling on equine Endothelial Colony Forming Cells (ECFCs) growth and function. It also aims at determining the duration of QD label retention and the mechanisms behind QD label loss. The motive behind this is to find a practical method of tracking stem and progenitor cells to monitor engraftment after administration, especially in the context of ischemic disease.

  • ECFCs from three adult horses were used for the study, with both QD-labeled and unlabeled ECFCs tested.
  • The researchers measured cell proliferation by comparing the number of cell doublings per day (NCD) and the population doubling time (PDT) in both labeled and unlabeled cells.
  • The function of the cells was assessed by uptake of acetylated low-density lipoprotein (DiO-Ac-LDL) and tubule formation on growth factor containing matrix.

Results

The study found no detrimental impact of QD labeling on cell growth and function. The data revealed that the NCD and PDT did not vary significantly between the unlabeled and the QD labeled cells.

  • Additionally, parameters like the uptake of DiO-Ac-LDL and tubule formation, indicative of cell function, showed no significant difference between unlabeled and QD labeled cells.
  • It was observed that the ECFCs retain their QD labeling over seven passages with both 5 nM and 20 nM label concentrations. However, a gradual reduction in label intensity was noticed over time.
  • The mechanism of QD label loss was determined to be due to cell division.

Conclusion

The researchers concluded that equine ECFCs could be effectively labeled with QD, and the QD concentrations up to 20 nM do not hinder cell growth or function. The study emphasizes that the loss of QD labels results from cell division. The researchers propose that the use of QD labeling could be a valuable method for tracing ECFCs engraftment in live applications, potentially driving advances in cell-based therapies for treating vascular diseases.

Cite This Article

APA
Winter RL, Seeto WJ, Tian Y, Caldwell FJ, Lipke EA, Wooldridge AA. (2018). Growth and function of equine endothelial colony forming cells labeled with semiconductor quantum dots. BMC Vet Res, 14(1), 247. https://doi.org/10.1186/s12917-018-1572-3

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 14
Issue: 1
Pages: 247
PII: 247

Researcher Affiliations

Winter, Randolph L
  • Department of Clinical Sciences, Auburn University, College of Veterinary Medicine, Auburn, AL, USA.
Seeto, Wen J
  • Department of Chemical Engineering, Auburn University, Auburn, AL, USA.
Tian, Yuan
  • Department of Chemical Engineering, Auburn University, Auburn, AL, USA.
Caldwell, Fred J
  • Department of Clinical Sciences, Auburn University, College of Veterinary Medicine, Auburn, AL, USA.
Lipke, Elizabeth A
  • Department of Chemical Engineering, Auburn University, Auburn, AL, USA.
Wooldridge, Anne A
  • Department of Clinical Sciences, Auburn University, College of Veterinary Medicine, Auburn, AL, USA. aaw0002@auburn.edu.

MeSH Terms

  • Animals
  • Cell Proliferation
  • Cells, Cultured
  • Endothelial Cells / cytology
  • Horses
  • Quantum Dots
  • Semiconductors
  • Staining and Labeling / methods
  • Stem Cells / cytology

Conflict of Interest Statement

ETHICS APPROVAL AND CONSENT TO PARTICIPATE: All procedures were approved by the University’s Animal Use and Care Committee. CONSENT FOR PUBLICATION: Not applicable. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER’S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Citations

This article has been cited 2 times.
  1. Winter RL, Tian Y, Caldwell FJ, Seeto WJ, Koehler JW, Pascoe DA, Fan S, Gaillard P, Lipke EA, Wooldridge AA. Cell engraftment, vascularization, and inflammation after treatment of equine distal limb wounds with endothelial colony forming cells encapsulated within hydrogel microspheres. BMC Vet Res 2020 Feb 4;16(1):43.
    doi: 10.1186/s12917-020-2269-ypubmed: 32019556google scholar: lookup
  2. Finding EJT, Faulkner A, Nash L, Wheeler-Jones CPD. Equine Endothelial Cells Show Pro-Angiogenic Behaviours in Response to Fibroblast Growth Factor 2 but Not Vascular Endothelial Growth Factor A. Int J Mol Sci 2024 May 30;25(11).
    doi: 10.3390/ijms25116017pubmed: 38892205google scholar: lookup
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