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

• Published in: Biotechnology journal 2024-02, Vol.19 (2), p.e2300484-n/a
• Main Authors: He, Jing, Wang, Zhangfan, Ao, Chunchun, Tu, Chengshu, Zhang, Yaqi, Chang, Cheng, Xiao, Cuihong, Xiang, E, Rao, Wei, Li, Changyong, Wu, Dongcheng
• Format: Article
• Language: English
• Subjects: Animals; Homo1; Humans; Mesenchymal Stem Cell Transplantation - methods; Mesenchymal Stem Cells - metabolism; pharmacokinetics; Rats; RT‐qPCR; UC‐MSCs; Umbilical Cord - metabolism
• ISSN: 1860-6768; 1860-7314
• DOI: 10.1002/biot.202300484

Background Owing 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. Methods We 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. Results We 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. Conclusions In 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. Graphical and Lay Summary Mesenchymal stem cells (MSCs) have demonstrated therapeutic potential in various refractory diseases. In this study, the authors established a highly sensitive and specific Homo1‐based real‐time quantitative PCR analysis method to quantify distribution of umbilical cord‐derived MSCs (UC‐MSCs) in vivo. This method poses as a novel approach for investigating the distribution and metabolism of human UC‐MSCs in vivo.