【Title】

In vivo tracking of PKH26-labeled endothelial progenitor cells after transplantation into rats with chronic liver injury

【Source】

Journal of Clinical Rehabilitative Tissue Engineering Research  September 3, 2009  Vol.13 No. 36，7011-7014

【Author】

Liu Feng, Liu Zhi-da, Wu Nan, Cong Xu, Fei Ran, Chen Hong-song, Wei Lai

【Setting】

Institute of Hepatology, People’s Hospital, Peking University, Beijing 100044, China

【Found】

Supported by: the National Youth Science Foundation of China, No. 30700350*; the National Key Basic Research and Development Plan of China, No. 2005CB522902, 2007CB512900**; the Foundation of People’s Hospital of Peking University, No. RDK2008-06*; the Science and Technology Plan of Beijing City, No. Z0006264040791*

【Abstract】

BACKGROUND: In our previous studies, we showed that endothelial progenitor cells (EPCs) ameliorate established liver fibrosis. But, many question, such as the way of EPCs infused, distribution of EPCs in injured liver tissue, need to be solved before EPCs were applied in the clinic.

OBJECTIVE: To track the PKH26-labeled EPCs after transplantation into the liver in rats with chronic liver injury.

DESIGN, TIME AND SETTING: The cytological in vivo study was performed at the Institute of Hepatology, People’s Hospital, Peking University from August 2008 to February 2009.

MATERIALS: A total of 100 healthy adult SPF grade Sprague Dawley rats were supplied by the Animal Center, Academy of Military Medical Sciences of Chinese PLA. Red fluorochrome PKH26 was purchased from Sigma, USA.

METHODS: EPCs were isolated from 16 rats by the adherence method, and then labeled with PKH26 in vitro. Three groups were set in this study. Four rats in the normal control group were left intact. Forty rats in the dimethylnitrosamine group were injected with 10 mg/kg dimethylnitrosamine by intraperitoneal injection to establish models of hepatic fibrosis. Forty rats in the carbon tetrachloride group were treated with carbon tetrachloride by gavage to generate models of hepatic fibrosis. Following model induction, rats in the dimethylnitrosamine and carbon tetrachloride groups were infused with 1 mL PKH26-labeled EPCs suspension (1×10⁶ cells).

MAIN OUTCOME MEASURES: PKH26 labeling rate and cell survival rate were measured. EPCs migration and CD31 expression were observed in rat liver under the fluorescence confocal microscopy.

RESULTS: Flow cytometry results demonstrated that positive rate of PKH26-labeled EPCs was 99%. Fluorescence microscopy showed that the survival rate of PKH26-labeled EPCs was >90%. One day following infusion, PKH26-labeled cells were observed in the liver of two forms of models of chronic hepatic injury, and the number of PKH26-labeled cells was increased over time. PKH26-labeled cells were mainly distributed in blood vessel endothelium along fibers and hepatic sinusoids in hepatic lobule. CD31/PKH26 double positive cells could be detected along vascular walls.

CONCLUSION: PKH26 could be used to label and track EPCs in the liver of rats with chronic liver injury in vitro.

【Number of pictures】

9 figures

【Full text】

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