title: Biological effects of iron oxide-protamine sulfate complex on mesenchymal stem cells and its relaxometry based labeling optimization for cellular MRI.
abstract: Mesenchymal stem cells (MSCs) are frequently used as a therapeutic, but reliable imaging technique to longitudinally evaluate the engraftment of transplanted cells is inadequate. For magnetic resonance imaging (MRI), it is essential to understand the technical competence of in vitro stem cells labeling with iron oxide with regard to its relaxation behavior and significance of its biological expressions. The purpose of the study was to optimize the effective labeling of MSCs with high transverse relaxivity iron oxide contrast agent with protamine sulfate and also evaluate the biological effects (phenotype and function) of labeled MSCs. Our results demonstrated that 50:3µg/ml of Fe-Pro complex containing 10% serum at an incubation time of 6h were ideal for effective in vitro labeling. Relaxometry study demonstrated that almost an 8-fold increase in relaxation rate (R2) was observed in labeled MSCs by comparing with unlabeled. Marginal alteration in Oct4 and CD146 genes, and phenotypic CD45 expressions were detected after labeling. T2-weighted images and histological analysis confirmed the homing of transplanted cells to the site of injury. The relaxometry based optimized labeling method of MSCs could be extrapolated for cellular MRI and may be useful in stem cell tracking in various pre-clinical and clinical studies.
authors.affiliation: NMR Research Centre, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, Delhi-54, India; Division of Stem Cell and Gene Therapy Research, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, Delhi-54, India.
authors.forename: Sushanta Kumar
authors.initials: SK
authors.lastname: Mishra
authors.affiliation: NMR Research Centre, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, Delhi-54, India. Electronic address: skhushu@yahoo.com.
authors.forename: Subash
authors.initials: S
authors.lastname: Khushu
authors.affiliation: Division of Stem Cell and Gene Therapy Research, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, Delhi-54, India. Electronic address: gugdutta@rediffmail.com.
authors.forename: Gurudutta
authors.initials: G
authors.lastname: Gangenahalli
mesh_terms: D000818:Animals; D051929:CD146 Antigen / Q000235:genetics / Q000378:metabolism; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D005290:Ferric Compounds / Q000009:adverse effects* / Q000737:chemistry; D017493:Leukocyte Common Antigens / Q000235:genetics / Q000378:metabolism; D008279:Magnetic Resonance Imaging / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011479:Protamines / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D051929:CD146 Antigen; D005290:Ferric Compounds; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011479:Protamines; D017493:Leukocyte Common Antigens
keywords: Fe-Pro complex; Magnetic resonance imaging; Mesenchymal stem cells; Relaxometry; Ultrasmall superparamagnetic iron oxide nanoparticles