|
|
||
 |
|||
|
|||
Originally published as JHC exPRESS on June 27, 2005.
doi:10.1369/jhc.5A6639.2005
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Volume 53 (10): 1215-1226, 2005
Copyright ©The Histochemical Society, Inc.
Analysis of Green Fluorescent Protein Expression in Transgenic Rats for Tracking Transplanted Neural Stem/Progenitor Cells
Division of Cellular and Molecular Biology, Toronto Western Research Institute, Toronto, Ontario, Canada (AJM,IK,RLvB,LL,CHT); Divisions of Organ Replacement Research, Animal Transgenic Research and Molecular Immunology, Centre for Molecular Medicine, Jichi Medical School, Kawachi, Tochigi, Japan (EK); and Princess Margaret Hospital, Ontario Cancer Institute, Toronto, Ontario, Canada (AK)
Correspondence to: Dr. Charles H. Tator, Toronto Western Hospital, McL 12-423, 399 Bathurst Street, Toronto, ON M5T 2S8, Canada. E-mail: charles.tator{at}uhn.on.ca
Green fluorescent protein (GFP) expression was evaluated in tissues of different transgenic rodents—Sprague-Dawley (SD) rat strain [SD-Tg(GFP)Bal], W rat strain [Wistar-TgN(CAG-GFP)184ys], and M mouse strain [Tg(GFPU)5Nagy/J]—by direct fluorescence of native GFP expression and by immunohistochemistry. The constitutively expressing GFP transgenic strains showed tissue-specific differences in GFP expression, and GFP immunohistochemistry amplified the fluorescent signal. The fluorescence of stem/progenitor cells cultured as neurospheres from the ependymal region of the adult spinal cord from the GFP SD and W rat strains was assessed in vitro. After transplantation of the cells into wild-type spinal cord, the ability to track the grafted cells was evaluated in vivo. Cultured stem/progenitor cells from the SD strain required GFP immunostaining to be visualized. Likewise, after transplantation of SD cells into the spinal cord, immunohistochemical amplification of the GFP signal was required for detection. In contrast, GFP expression of stem/progenitor cells generated from the W strain was readily detected by direct fluorescence both in vitro and in vivo without the need for immunohistochemical amplification. The cultured stem/progenitor cells transplanted into the spinal cord survived for at least 49 days after transplantation, and continued to express GFP, demonstrating stable expression of the GFP transgene in vivo. (J Histochem Cytochem 53:1215–1226, 2005)
Key Words: GFP • transgenic rat • immunohistochemistry • spinal cord • stem/progenitor cells • transplantation
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
 |
|
  I. Kulbatski and C. H. Tator Region-specific Differentiation Potential of Adult Rat Spinal Cord Neural Stem/Precursors and Their Plasticity in Response to In Vitro Manipulation J. Histochem. Cytochem., May 1, 2009; 57(5): 405 - 423. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. Coppen, S. Fukushima, Y. Shintani, K. Takahashi, A. Varela-Carver, H. Salem, K. Yashiro, M. H. Yacoub, and K. Suzuki A Factor Underlying Late-Phase Arrhythmogenicity After Cell Therapy to the Heart: Global Downregulation of Connexin43 in the Host Myocardium After Skeletal Myoblast Transplantation Circulation, September 30, 2008; 118(14_suppl_1): S138 - S144. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Kulbatski, A. J. Mothe, A. Keating, Y. Hakamata, E. Kobayashi, and C. H. Tator Oligodendrocytes and Radial Glia Derived From Adult Rat Spinal Cord Progenitors: Morphological and Immunocytochemical Characterization J. Histochem. Cytochem., March 1, 2007; 55(3): 209 - 222. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Hannouche, A. Raould, R.S. Nizard, L. Sedel, and H. Petite Embedding of Bone Samples in Methylmethacrylate: A Suitable Method for Tracking LacZ Mesenchymal Stem Cells in Skeletal Tissues J. Histochem. Cytochem., March 1, 2007; 55(3): 255 - 262. [Abstract] [Full Text] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |
