Methods to assess stem cell lineage, fate and function

Nguyen, Patricia K.; Nag, Divya; Wu, Joseph C.

doi:10.1016/j.addr.2010.08.008

Cited by 45 publications

(34 citation statements)

References 80 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A range of technologies that are available include those adapted from the radiologic setting and others based on optical techniques that build on the methods of optical microscopy and diffuse optical tomography [25]. A powerful approach is to use imaging reporter genes or direct radio-labeling in conjunction with these imaging technologies [11, 12, 14]. The most common reporter system used has been the gene for firefly luciferase and its substrate luciferin for BLI for experimental studies.…”

Section: Discussionmentioning

confidence: 99%

Radiolabeling and In Vivo Imaging of Transplanted Renal Lineages Differentiated from Human Embryonic Stem Cells in Fetal Rhesus Monkeys

et al. 2011

View full text Add to dashboard Cite

Purpose The goals of this study were to optimize radiolabeling of renal lineages differentiated from human embryonic stem (hES) cells and use noninvasive imaging (positron emission tomography (PET) and bioluminescence imaging (BLI)) to detect the cells in fetal monkeys post-transplant. Procedures hES cells expressing firefly luciferase (5×106) were radiolabeled with the optimized concentration of 10 μCi/ml 64Cu-PTSM then transplanted under ultrasound guidance into early second trimester fetal monkey kidneys. Fetuses were imaged in utero with PET and tissues collected for analysis 3 days post-transplant. Fetal kidneys were imaged ex vivo (PET and BLI) post-tissue harvest, and serial kidney sections were assessed by PCR for human-specific DNA sequences, fluorescent in situ hybridization (FISH) for human-specific centromere probes, and immunohistochemistry (IHC) to assess engrafted cells. Results Transplanted cells were readily imaged in vivo and identified at the site of injection; tissue analyses confirmed the imaging findings. Using a semi-quantitative method, one in approximately 650 cells in the kidney was shown to be of human origin by PCR and FISH. Conclusions These studies suggest that hES cells differentiated toward renal lineages can be effectively radiolabeled, transplanted into fetal monkey kidneys under ultrasound guidance, monitored with PET post-transplant, and identified by PET, BLI, PCR, FISH, and IHC post-tissue harvest.

show abstract

Section: Discussionmentioning

confidence: 99%

Radiolabeling and In Vivo Imaging of Transplanted Renal Lineages Differentiated from Human Embryonic Stem Cells in Fetal Rhesus Monkeys

et al. 2011

View full text Add to dashboard Cite

show abstract

“…Following administration, depending on the route as many as 90 % of transplanted stem cells may be lost due to a combination of physical stress, infl ammation, hypoxia, cell death following detachment, or immunogenic rejection (Nguyen et al 2010 ;Zvibel et al 2002 ). The half-life of transplanted cells is an important consideration-a short survival time may require multiple treatments for a chronic indication or delivery of a large number of stem cells.…”

Section: Pharmacokinetics and Pharmacodistribution (Pk/pd)mentioning

confidence: 99%

“…Although treatments are often administered to immune-competent human patients , especially in cases of allogeneic cell therapies involving immunosupression, the tumorigenic potential of a small set of pluripotent stem cells merits consideration. Unfortunately, no current analytical technique is suffi ciently sensitive to assure removal of all pluripotent cells (Nguyen et al 2010 ). As an additional safety measure, some are considering incorporating suicide genes or stem cell specifi c toxins to mitigate the risk of tumors from malignant stem cell growth (Knoepfl er 2009 ).…”

Section: Tumorigenicitymentioning

confidence: 99%

Guidelines for Preclinical Development

Spack

2016

Regenerative Medicine - From Protocol to Patient

View full text Add to dashboard Cite

Preclinical development encompasses the set of activities required to initiate testing for safety and effi cacy in humans. These requirements are defi ned by national and international regulations, and are generally focused on defi nition of drug composition and safety as well as scientifi c rationale and establishing a proposed dose regimen. The preclinical regulatory requirements for cell therapies broadly parallel those for small molecules and proteins, but the complexity of cells can bring additional challenges to the defi nition of potency, composition, immunogenicity, and toxicity. As a consequence of their pluripotent origins, stem cell therapies require additional preclinical considerations related to potential tumorigenicity and genetic/epigenetic stability. The fi rst FDA approved stem cell therapy study initiated clinical trial in 2010. Thus, the fi eld is young and there are few completed studies to provide precedents for planned preclinical development of novel stem cell approaches. Rather, the fi eld must draw practical lessons from other somatic cell therapies. The evolution and harmonization of regulatory guidances for stem cells must also contend with a growing number of unregulated sites around the world that offer unproven stem cell treatments. It takes time for any new technology to identify safety and effi cacy barriers to clinical application, and to develop strategies to overcome them. As current research and clinical experience guide the next generation of stem cell therapies in preclinical development, the establishment of validated assays based on cells differentiated from human stem cells may revolutionize the preclinical safety testing for all classes of drugs.

show abstract

“…Labeling cells with iron particles for MRI visualization is one of the most frequently applied methods for cell tracking. The most commonly used iron formulation is superparamagnetic iron oxide particles because of their potent negative contrast effects and inherent lack of cell toxicity (56). The labeling of iron particles has to be completed ex vivo before transplantation, because most stem cells must be induced to take up these MRI contrast agents (28).…”

Section: Molecular Imaging For the Survival And Kinetics Of Engraftedmentioning

confidence: 99%

Mechanistic molecular imaging of cardiac cell therapy for ischemic heart disease

Fan

Cao

2013

American Journal of Physiology-Heart and Circulatory Physiology

View full text Add to dashboard Cite

Yu Q, Fan W, Cao F. Mechanistic molecular imaging of cardiac cell therapy for ischemic heart disease. Am J Physiol Heart Circ Physiol 305: H947-H959, 2013. First published July 26, 2013 doi:10.1152/ajpheart.00092.2013.-Cell-based myocardial regeneration has emerged as a promising therapeutic option for ischemic heart disease, though not yet at the level of routine clinical utility. Despite the encouraging results from initial preclinical studies that have demonstrated improved function and reduced infarct size of the ischemic myocardium following several candidate cell transplantation, the beneficial effects and molecular mechanisms of cardiac cell therapy are still unclear in clinical applications to date, and much remains to be optimized. To improve engraftment, accurate methods are required for tracking cell fate and quantifying functional outcome. In the present review, we summarized the current status and challenges of cardiac cell therapy for ischemic heart disease and discussed the strengths and limitations of currently available in vivo imaging techniques with special focus on the newly developed multimodality approaches for assessing the efficacy of engrafted donor cells. We also addressed the hurdles these imaging modalities are facing, including issues regarding immunogenicity and tumorigenicity of transplanted stem cells, and provided some the future perspectives on stem cell imaging. molecular imaging; stem cell therapy; ischemic heart disease THIS ARTICLE is part of a collection on Physiological Basis of Cardiovascular Cell and Gene Therapies. Other articles appearing in this collection, as well as a full archive of all collections, can be found online at http://ajpheart.physiology. org/.

show abstract

Methods to assess stem cell lineage, fate and function

Cited by 45 publications

References 80 publications

Radiolabeling and In Vivo Imaging of Transplanted Renal Lineages Differentiated from Human Embryonic Stem Cells in Fetal Rhesus Monkeys

Radiolabeling and In Vivo Imaging of Transplanted Renal Lineages Differentiated from Human Embryonic Stem Cells in Fetal Rhesus Monkeys

Guidelines for Preclinical Development

Mechanistic molecular imaging of cardiac cell therapy for ischemic heart disease

Contact Info

Product

Resources

About