Labeling and tracking of mesenchymal stromal cells with EdU

Guiting Lin, Yun Ching Huang, Alan W. Shindel, Lia Banie, Guifang Wang, Tom F. Lue, Ching Shwun Lin

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Background aims The thymidine analog bromodeoxyuridine (5-bromo-2- deoxyuridine; BrdU) has been used widely to label cells in culture and in tissue. The labeled cells can also be tracked when transplanted into a suitable host. In the present study we tested a new thymidine analog, 5-ethynyl-2-deoxyuridine (EdU), for labeling and tracking of mesenchymal stromal cells (MSC), specifically adipose tissue-derived stem cells (ADSC). Methods Labeling of ADSC was examined for the dosage effect of EdU and stability of label by Alexa-594 staining followed by fluorescence microscopy. Labeling of various organs/tissues was done by intraperitoneal injection of EdU and examined by histology and fluorescence microscopy. Tracking of ADSC was done by intratissue or intravenous transplantation of EdU-labeled ADSC into various tissues and examined by histology and fluorescence microscopy. Results EdU was incorporated specifically into the nucleus in approximately 50% of ADSC and the percentage of cells that remained fully labeled declined with time. Peritoneal injection of EdU resulted in the appearance of EdU-positive cells in most organs and tissues. In the intestine, EdU-positive cells were found in both the epithelium and connective tissues 7 h after injection. Long-term (26 week) follow-ups found EdU-positive cells only in the connective tissue. Tracking of ADSC was successful in tissues 10 weeks after intratissue or intravenous transplantation. Conclusions Cell labeling with EdU in culture or living animals can be performed easily. The detection of EdU label requires no harsh treatment or immunologic reaction, as detection of BrdU label does. EdU can be used for long-term tracking of ADSC.

Original languageEnglish (US)
Pages (from-to)864-873
Number of pages10
JournalCytotherapy
Volume11
Issue number7
DOIs
StatePublished - 2009

Fingerprint

Mesenchymal Stromal Cells
Adipose Tissue
Stem Cells
Bromodeoxyuridine
Fluorescence Microscopy
Connective Tissue
Thymidine
5-ethynyl-2'-deoxyuridine
Histology
Transplantation
Injections
Intraperitoneal Injections
Intestines
Epithelium
Cell Culture Techniques
Staining and Labeling

Keywords

  • Adipose tissue-derived stem cells
  • Cell labeling
  • Cell tracking
  • EdU
  • Mesenchymal stromal cells

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Molecular Medicine

Cite this

Lin, G., Huang, Y. C., Shindel, A. W., Banie, L., Wang, G., Lue, T. F., & Lin, C. S. (2009). Labeling and tracking of mesenchymal stromal cells with EdU. Cytotherapy, 11(7), 864-873. https://doi.org/10.3109/14653240903180084

Labeling and tracking of mesenchymal stromal cells with EdU. / Lin, Guiting; Huang, Yun Ching; Shindel, Alan W.; Banie, Lia; Wang, Guifang; Lue, Tom F.; Lin, Ching Shwun.

In: Cytotherapy, Vol. 11, No. 7, 2009, p. 864-873.

Research output: Contribution to journalArticle

Lin, G, Huang, YC, Shindel, AW, Banie, L, Wang, G, Lue, TF & Lin, CS 2009, 'Labeling and tracking of mesenchymal stromal cells with EdU', Cytotherapy, vol. 11, no. 7, pp. 864-873. https://doi.org/10.3109/14653240903180084
Lin, Guiting ; Huang, Yun Ching ; Shindel, Alan W. ; Banie, Lia ; Wang, Guifang ; Lue, Tom F. ; Lin, Ching Shwun. / Labeling and tracking of mesenchymal stromal cells with EdU. In: Cytotherapy. 2009 ; Vol. 11, No. 7. pp. 864-873.
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abstract = "Background aims The thymidine analog bromodeoxyuridine (5-bromo-2- deoxyuridine; BrdU) has been used widely to label cells in culture and in tissue. The labeled cells can also be tracked when transplanted into a suitable host. In the present study we tested a new thymidine analog, 5-ethynyl-2-deoxyuridine (EdU), for labeling and tracking of mesenchymal stromal cells (MSC), specifically adipose tissue-derived stem cells (ADSC). Methods Labeling of ADSC was examined for the dosage effect of EdU and stability of label by Alexa-594 staining followed by fluorescence microscopy. Labeling of various organs/tissues was done by intraperitoneal injection of EdU and examined by histology and fluorescence microscopy. Tracking of ADSC was done by intratissue or intravenous transplantation of EdU-labeled ADSC into various tissues and examined by histology and fluorescence microscopy. Results EdU was incorporated specifically into the nucleus in approximately 50{\%} of ADSC and the percentage of cells that remained fully labeled declined with time. Peritoneal injection of EdU resulted in the appearance of EdU-positive cells in most organs and tissues. In the intestine, EdU-positive cells were found in both the epithelium and connective tissues 7 h after injection. Long-term (26 week) follow-ups found EdU-positive cells only in the connective tissue. Tracking of ADSC was successful in tissues 10 weeks after intratissue or intravenous transplantation. Conclusions Cell labeling with EdU in culture or living animals can be performed easily. The detection of EdU label requires no harsh treatment or immunologic reaction, as detection of BrdU label does. EdU can be used for long-term tracking of ADSC.",
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T1 - Labeling and tracking of mesenchymal stromal cells with EdU

AU - Lin, Guiting

AU - Huang, Yun Ching

AU - Shindel, Alan W.

AU - Banie, Lia

AU - Wang, Guifang

AU - Lue, Tom F.

AU - Lin, Ching Shwun

PY - 2009

Y1 - 2009

N2 - Background aims The thymidine analog bromodeoxyuridine (5-bromo-2- deoxyuridine; BrdU) has been used widely to label cells in culture and in tissue. The labeled cells can also be tracked when transplanted into a suitable host. In the present study we tested a new thymidine analog, 5-ethynyl-2-deoxyuridine (EdU), for labeling and tracking of mesenchymal stromal cells (MSC), specifically adipose tissue-derived stem cells (ADSC). Methods Labeling of ADSC was examined for the dosage effect of EdU and stability of label by Alexa-594 staining followed by fluorescence microscopy. Labeling of various organs/tissues was done by intraperitoneal injection of EdU and examined by histology and fluorescence microscopy. Tracking of ADSC was done by intratissue or intravenous transplantation of EdU-labeled ADSC into various tissues and examined by histology and fluorescence microscopy. Results EdU was incorporated specifically into the nucleus in approximately 50% of ADSC and the percentage of cells that remained fully labeled declined with time. Peritoneal injection of EdU resulted in the appearance of EdU-positive cells in most organs and tissues. In the intestine, EdU-positive cells were found in both the epithelium and connective tissues 7 h after injection. Long-term (26 week) follow-ups found EdU-positive cells only in the connective tissue. Tracking of ADSC was successful in tissues 10 weeks after intratissue or intravenous transplantation. Conclusions Cell labeling with EdU in culture or living animals can be performed easily. The detection of EdU label requires no harsh treatment or immunologic reaction, as detection of BrdU label does. EdU can be used for long-term tracking of ADSC.

AB - Background aims The thymidine analog bromodeoxyuridine (5-bromo-2- deoxyuridine; BrdU) has been used widely to label cells in culture and in tissue. The labeled cells can also be tracked when transplanted into a suitable host. In the present study we tested a new thymidine analog, 5-ethynyl-2-deoxyuridine (EdU), for labeling and tracking of mesenchymal stromal cells (MSC), specifically adipose tissue-derived stem cells (ADSC). Methods Labeling of ADSC was examined for the dosage effect of EdU and stability of label by Alexa-594 staining followed by fluorescence microscopy. Labeling of various organs/tissues was done by intraperitoneal injection of EdU and examined by histology and fluorescence microscopy. Tracking of ADSC was done by intratissue or intravenous transplantation of EdU-labeled ADSC into various tissues and examined by histology and fluorescence microscopy. Results EdU was incorporated specifically into the nucleus in approximately 50% of ADSC and the percentage of cells that remained fully labeled declined with time. Peritoneal injection of EdU resulted in the appearance of EdU-positive cells in most organs and tissues. In the intestine, EdU-positive cells were found in both the epithelium and connective tissues 7 h after injection. Long-term (26 week) follow-ups found EdU-positive cells only in the connective tissue. Tracking of ADSC was successful in tissues 10 weeks after intratissue or intravenous transplantation. Conclusions Cell labeling with EdU in culture or living animals can be performed easily. The detection of EdU label requires no harsh treatment or immunologic reaction, as detection of BrdU label does. EdU can be used for long-term tracking of ADSC.

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KW - Cell tracking

KW - EdU

KW - Mesenchymal stromal cells

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