Assessment of biological characteristics of adipose tissue-derived stem cells co-labeled with Molday ION Rhodamine B™ and green fluorescent protein in vitro.
The current study aimed to investigate adipose tissue-derived stem cells (ADSCs) in vivo by multimodality imaging following implantation for cellular therapy. The biological characteristics of ADSCs co-labeled with Molday ION Rhodamine B™ (MIRB) and green fluorescent protein (GFP) were studied in vitro. Following rat ADSC isolation and culture, a combined labeling strategy for ADSCs based on genetic modification of the reporter gene GFP with lentiviral vector expression enhancement and physical MIRB labeling was performed. Cell viability, proliferation, membrane-bound antigens and multiple differentiation ability were compared between the labeled and unlabeled ADSCs. The ADSCs were successfully labeled with GFP and MIRB, showing various fluorescent colors for marker identification. The fluorescence emitted by the GFP protein was sustained and exhibited stable expression, while MIRB fluorescence decreased with time. Compared with the unlabeled ADSCs, no significant differences were detected in cell viability, proliferation, membrane-bound antigens and multiple differentiation ability in the co-labeled samples (P>0.05). No significant effects on the biophysical properties of ADSCs were observed following co-labeling with lentiviral vectors encoding the gene for emerald green fluorescent protein and MIRB. The ADSCs were able to be efficiently tracked in vitro and in vivo by multimodality imaging thus, the co-labeling approach provides a novel strategy for therapeutic gene studies.
Plastic Surgery Institute, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510282, P.R. China.
Hua
H
Nan
Jiacheng
J
Huang
Hongmian
H
Li
Qiong
Q
Li
Dalie
D
Liu
D000273:Adipose Tissue / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002468:Cell Physiological Phenomena / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D052203:Ferrosoferric Oxide / Q000493:pharmacokinetics*; D005434:Flow Cytometry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D058185:Magnetite Nanoparticles / Q000737:chemistry; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012235:Rhodamines / Q000493:pharmacokinetics*; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014018:Tissue Distribution
D005456:Fluorescent Dyes; D058185:Magnetite Nanoparticles; D012235:Rhodamines; D049452:Green Fluorescent Proteins; D052203:Ferrosoferric Oxide