Abstract
Cell therapy is a promising strategy for the treatment of human diseases. While the first use of cells for therapeutic purposes can be traced to the 19th century, there has been a lack of general and reliable methods to study the biodistribution and associated pharmacokinetics of transplanted cells in various animal models for preclinical evaluation. Here, we present a new platform using albumin-conjugated fluorescent nanodiamonds (FNDs) as biocompatible and photostable labels for quantitative tracking of human placenta choriodecidual membrane-derived mesenchymal stem cells (pcMSCs) in miniature pigs by magnetic modulation. With this background-free detection technique and time-gated fluorescence imaging, we have been able to precisely determine the numbers as well as positions of the transplanted FND-labeled pcMSCs in organs and tissues of the miniature pigs after intravenous administration. The method is applicable to single-cell imaging and quantitative tracking of human stem/progenitor cells in rodents and other animal models as well.
Highlights
Particular, when applied to large animals such as pigs, which share many physiological similarities with humans[9], the PCR methods often yield unreliable results (Supplementary Table S1)
The key instrument used in this quantification is a home-built magnetically modulated fluorescence (MMF) spectrometer (Supplementary Fig. S1)
We have developed MMF into an innovative method for absolute and sensitive quantification of fluorescent nanodiamond (FND) in biological matrices
Summary
Particular, when applied to large animals such as pigs, which share many physiological similarities with humans[9], the PCR methods often yield unreliable results (Supplementary Table S1). The particle contains a high-density ensemble of negatively charged nitrogen-vacancy (NV−) centers as built-in fluorophores, which are red fluorescent[17], perfectly photostable[18], and can be detected with high sensitivity in cells and tissues by fluorescence time gating[19], microwave modulation[20,21], and magnetic modulation[22]. These distinctive characteristics together make it possible to follow the biodistribution of human MSCs after FND labeling and subsequent transplantation in animals such as miniature pigs. Engraftment of the stem cells in tissues was identified by time-gated fluorescence imaging with single-cell resolution[19]
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