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J Invest Surg. 2012 Dec;25(6):405-14. doi: 10.3109/08941939.2012.661519.

A technique for systemic mesenchymal stem cell transplantation in newborn rat pups.

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Department of Pediatric Surgery, College of Medicine, The Research Institute at Nationwide Children's Hospital, Center for Perinatal Research, Nationwide Children's Hospital, The Ohio State University, Columbus, OH 43205, USA.


Mesenchymal stem cells (MSC) have the potential to aid tissue regeneration. Intravenous (IV) MSC administration is currently being assessed following tissue injury. However, few studies have been performed to establish a safe and effective method of IV MSC infusion for newborns. We have established a safe, nontraumatic and effective technique for systemic MSC transplantation in newborn rats. Yellow-fluorescent-protein (YFP)-labeled MSC were characterized using MSC markers and their differentiation potential was confirmed. Rat pups were delivered by C-section on gestational day 21. The umbilical vein (UV) was cannulated and used for IV injection of MSC or saline control, which was performed under ultrasonographic imaging. An additional control group consisted of UV MSC injection in adult mice. Mean operating time, success rate of cannulation and death rate were recorded. YFP-MSC quantification in multiple organs was performed. Mean operating time was 3.9 ± 1.1 min. The success of UV MSC injection was 92.8%. The immediate and 24 hr delayed death rate for rat pups was significantly lower than that of adult mice (p < .05). No pups receiving saline injection died. After locating the patent foramen ovale (PFO) of newborn pups by ultrasonographic imaging, extra pulse-waves and wave-shape changes were detected when MSC were injected. The number of YFP-MSC was 15.8 ± 4.1 cells per visual field (CPVF) in the lungs, 2.9 ± 1.2 CPVF in the heart, and 19.8 ± 5.0 CPVF in the intestines. We conclude that IV MSC infusion through the UV is a convenient, safe, and effective method for systemic MSC transplantation in prematurely delivered newborn rats.

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