Cytotechnology

, Volume 64, Issue 4, pp 391–401

Assessment of the green florescence protein labeling method for tracking implanted mesenchymal stem cells

  • Yinghua Guo
  • Longxiang Su
  • Junlou Wu
  • Dong Zhang
  • Xiaojun Zhang
  • Guizhi Zhang
  • Tianzhi Li
  • Junfeng Wang
  • Changting Liu
Original Research

DOI: 10.1007/s10616-011-9417-y

Cite this article as:
Guo, Y., Su, L., Wu, J. et al. Cytotechnology (2012) 64: 391. doi:10.1007/s10616-011-9417-y

Abstract

Although green fluorescent protein (GFP) labeling is widely accepted as a tracking method, much remains uncertain regarding the retention of injected GFP-labeled cells implanted in ischemic organs. In this study, we evaluate the effectiveness of GFP for identifying and tracking implanted bone marrow- mesenchymal stem cells (BM-MSCs) and the effect of GFP on the paracrine actions of these cells. MSCs isolated from rat femur marrow were transduced with a recombinant adenovirus carrying GFP. After transplantation of the GFP-labeled BM-MSCs into the infarct zone of rat hearts, the survival, distribution, and migration of the labeled cells were analyzed at 3, 7, 14, and 28 days. To evaluate the effect of GFP on the paracrine actions of BM-MSCs, Western blot analysis was performed to detect the expression of vascular endothelial growth factor (VEGF), b fibroblast growth factor (b FGF), tissue inhibitor of metalloproteinase-1 (TIMP-1) and matrix metalloproteinases-2 (MMP-2). GFP was successfully expressed by BM-MSCs in vitro. At 14 days after cell transplantation the GFP-positive cells could not be detected via confocal microscopy. By using a GFP antibody, distinct GFP-positive cells could be seen and quantitative analysis showed that the expression volume of GFP was 6.42 ± 0.92 mm3 after 3 days, 1.24 ± 0.76 mm3 after 7 days, 0.33 ± 0.03 mm3 after 14 days, and 0.09 ± 0.05 mm3 after 28 days. GFP labeling did not adversely affect the paracrine actions of BM-MSCs. GFP labeling could be used to track MSC distribution and their fate for at least 28 days after delivery to rat hearts with myocardial infarction, and this stem cell tracking strategy did not adversely affect the paracrine actions of BM-MSCs.

Keywords

Cell tracking Green fluorescent protein Mesenchymal stem cells paracrine Myocardial infarction 

Abbreviations

GFP

Green fluorescent protein

BM-MSCs

Bone marrow-mesenchymal stem cells

SC

Stem cell

Ad-GFP

Adenovirus vector carrying GFP

SD rats

Sprague–Dawley rats

MI

Myocardial infarction

MOIs

Multiplicities of infection

PBS

Phosphate-buffered saline

LV

Left ventricle

VEGF

Vascular endothelial growth factor

FGF

Fibroblast growth factor

TIMP-1

Tissue inhibitor of metalloproteinase-1

MMP-2

Matrix metalloproteinases-2

vWF

von Willebrand factor

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Yinghua Guo
    • 1
  • Longxiang Su
    • 1
  • Junlou Wu
    • 2
  • Dong Zhang
    • 1
  • Xiaojun Zhang
    • 1
  • Guizhi Zhang
    • 1
  • Tianzhi Li
    • 1
  • Junfeng Wang
    • 1
  • Changting Liu
    • 1
  1. 1.Department of Nanlou Respirtory DiseaseChinese PLA General Hospital, Chinese PLA Postgraduate Medical SchoolBeijingPeople’s Republic of China
  2. 2.From Shandong Provincial HospitalShandong University School of MedicineJinanChina

Personalised recommendations