A highly sensitive and specific Homo1‐based real‐time qPCR method for quantification of human umbilical cord mesenchymal stem cells in rats

Author:

He Jing1,Wang Zhangfan2,Ao Chunchun1,Tu Chengshu3,Zhang Yaqi1,Chang Cheng1,Xiao Cuihong2,Xiang E2,Rao Wei2,Li Changyong45ORCID,Wu Dongcheng126

Affiliation:

1. Department of Biochemistry and Molecular Biology Wuhan University School of Basic Medical Sciences Wuhan China

2. R&D Center Wuhan Hamilton Biotechnology Co. Ltd Wuhan China

3. Department of Oncology Nanfang Hospital Southern Medical University Guangzhou China

4. Department of Physiology Wuhan University School of Basic Medical Sciences Wuhan China

5. Xianning Medical College Hubei University of Science & Technology Xianning China

6. R&D Center Guangzhou Hamilton Biotechnology Co. Ltd Guangzhou China

Abstract

AbstractBackgroundOwing to the characteristics of easier access in vitro, low immunogenicity, and high plasticity, human umbilical cord‐derived mesenchymal stem cells (UC‐MSCs) are considered as a promising cell‐based drugs for clinical application. No internationally recognized technology exists to evaluate the pharmacokinetics and distribution of cell‐based drugs in vivo.MethodsWe determined the human‐specific gene sequence, Homo1, from differential fragments Homo sapiens mitochondrion and Rattus norvegicus mitochondrion. The expression of Homo1 was utilized to determine the distribution of UC‐MSCs in the normal and diabetic nephropathy (DN) rats.ResultsWe observed a significant correlation between the number of UC‐MSCs and the expression level of Homo1. Following intravenous transplantation, the blood levels of UC‐MSCs peaked at 30 min. A large amount of intravenously injected MSCs were trapped in the lungs, but the number of them decreased rapidly after 24 h. Additionally, the distribution of UC‐MSCs in the kidneys of DN rats was significantly higher than that of normal rats.ConclusionsIn this study, we establish a highly sensitive and specific Homo1‐based real‐time quantitative PCR method to quantify the distribution of human UC‐MSCs in rats. The method provides guidelines for the safety research of cells in preclinical stages.

Funder

Wuhan Municipal Science and Technology Bureau

Publisher

Wiley

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