Cellular Iron Metabolism Studies Demonstate Safety of Magnetic Tracking of Mesenchymal Stem Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4320-4320
Author(s):  
Edyta Pawelczyk ◽  
Ali S. Arbab ◽  
Sunil Pundit ◽  
Joseph A. Frank
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Light Microscopy ◽  
Iron Metabolism ◽  
Short Term ◽  
Protein Levels ◽  
Cellular Iron ◽  
Magnetic Labeling ◽  
And Storage

Abstract Background: Ferumoxides-protamine sulfate (Fe-Pro) complexes are used for intracellular magnetic labeling of mesenchymal stem cells (MSCs) to non invasively monitor cell trafficking and tissue distribution by in vivo magnetic resonance imaging (MRI). It has been previously shown that there was no short or long- term toxic effects on cell’s viability, proliferation, reactive oxygen species and biological functions including differentiation capacity of MSCs, however the effects of dextran coated superparamagnetic iron oxide (SPIO) nanoparticles labeling on cellular iron metabolism and storage is unknown. Objective: Evaluate the effect of using Fe-Pro complexes for magnetic cell labeling in MSCs on the expression of transferrin receptor (TfR-1) and ferritin, proteins involved in intracellular iron metabolism and storage. Methods: Confluent MSCs were labeled with Fe-Pro complexes at the ratio of 100 ug/mL to 6 ug/mL. After labeling, cells were washed and cultured for 2 months. TfR-1 and ferritin expression were evaluated at the gene and protein levels in total cell lysates, on day 1, 3, 7, 14, 28 and 35 days post-labeling, by real-time polymerase chain reaction (RT-PCR) and Western blotting. MSCs were evaluated for the presence of iron by Prussian blue stain and light microscopy. In addition, TfR-1 and ferritin levels were determined for Fe-Pro labeled and unlabeled MSCs, allowed to slowly multiply, as a result of dividing the cells in half when they reached confluence. Results: All cells were Prussian blue positive on light microscopy following labeling with Fe-Pro. Labeling of MSCs grown to confluence resulted in short-term increase of TfR-1 mRNA (day 1 and day 3, p= 0.0056) without changes in TfR-1 protein levels. Fe- Pro labeled MSC demonstrated an increase in ferritin gene expression on day 7 and 14 (p=0.0003) compared to unlabeled cells, while protein levels were higher compared to unlabeled MSC at each time point (p=0.005). However, ferritin protein levels from Fe-Pro labeled MSCs were not significantly increased after 7 passages. Conclusions: Magnetic labeling of cells with FDA approved ferumoxides complexed to protamine sulfate elicited short-term changes in cellular iron metabolism and storage that was dependent on ability to divide. Fe-Pro labeling did not have any long-term implications validating the safety of Fe-Pro labeling technique for MSCs trafficking studies by MRI.

scholarly journals Long-Term Sevoflurane Exposure Inhibits The Proliferation, Differentiation, and Homing Potential of Bone Marrow Mesenchymal Stem Cells

2021 ◽  
Author(s):  
Yunxiao Bai ◽  
Peng Wang ◽  
Xiaofang Ma ◽  
Lili Li ◽  
Chunyan Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Wound Repair ◽  
Vascular Endothelial Cells ◽  
Short Term ◽  
Repair Gene ◽  
Anesthetic Drugs ◽  
Marrow Mesenchymal Stem Cells

Abstract Background: Bone marrow mesenchymal stem cells (BMSCs) are widely used in many fields such as wound repair, gene delivery, and microenvironment improvement. In some cases, BMSC transplantation requires long-term anesthesia. However, the effects of anesthetics on the characteristics of BMSCs are poorly understood.Methods: In this study, we examined the effect of sevoflurane, a gas anesthetic drug most commonly used in children, on the proliferation, differentiation, and homing potential of BMSCs.Results: Short-term (6 h) sevoflurane exposure had almost no effect on the proliferation, differentiation, and homing of BMSCs. However, long-term (24 h) sevoflurane exposure inhibited the proliferation of BMSCs, accelerated their differentiation into nerve cells, and inhibited their homing potential to damaged vascular endothelial cells and intact glioma cells.Conclusion: Short-term anesthesia with sevoflurane as the main inducer is safe and harmless to BMSCs, but long-term sevoflurane exposure may reduce their repair potential. Therefore, because of the high proportion of BMSCs in children, the application of long-term anesthesia with sevoflurane should be cautious, or more suitable anesthetic drugs are needed.

scholarly journals Dry storage of sperm: applications in primates and domestic animals

2006 ◽  
Vol 18 (2) ◽  
pp. 1 ◽  
Author(s):  
Stuart A. Meyers
Keyword(s):  
Stem Cells ◽  
Spermatogonial Stem Cells ◽  
Short Term ◽  
Long Term Storage ◽  
Space Requirements ◽  
Thermotropic Phase Transitions ◽  
And Storage ◽  
Cryogenic Conditions ◽  
Term Storage

Cryopreservation of spermatozoa, oocytes and embryos, as well as somatic cells or cell lines for cloning from cells, are all options for the long-term storage of unique genotypes and endangered species. Spermatozoal cryopreservation and storage currently require liquid nitrogen or ultralow refrigeration-based methods for long- or short-term storage, which requires routine maintenance and extensive space requirements. The preservation of stem cells also has strict requirements for long-term storage to maintain genetic integrity. Dessicated (lyopreserved) sperm and stem cells will provide an unprecedented type of long-term storage without the need for expensive and burdensome cryogenic conditions. Experiments were conducted to determine an effective intracellular concentration of the lyoprotectant trehalose. High-pressure liquid chromatography studies revealed that trehalose can be incorporated into mature sperm cells as well as spermatogonial stem cells from rhesus monkeys. In addition, using fourier transform infrared spectroscopy, we determined that thermotropic phase transitions for fresh ejaculates from rhesus monkey and stallion sperm occurred at 10–15, 33–37 and 55–59°C. Preliminary studies in our laboratory have indicated that spermatogonial stem cells can be dried to <3 g g−1 water and maintain viability following rehydration. Studies in our laboratory have provided preliminary results suggesting that the desiccated storage of sperm and spermatogonial stem cells may be a viable alternative to conventional cryopreservation.

scholarly journals Senescence-Associated Secretory Phenotype Suppression Mediated by Small-Sized Mesenchymal Stem Cells Delays Cellular Senescence through TLR2 and TLR5 Signaling

Cells ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 63
Author(s):  
Ji Hye Kwon ◽  
Miyeon Kim ◽  
Soyoun Um ◽  
Hyang Ju Lee ◽  
Yun Kyung Bae ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cellular Senescence ◽  
Small Cells ◽  
Critical Determinant ◽  
Senescent Phenotype ◽  
Toll Like Receptor 2 ◽  
Secretory Phenotype ◽  
Major Factors

In order to provide a sufficient number of cells for clinical use, mesenchymal stem cells (MSCs) must be cultured for long-term expansion, which inevitably triggers cellular senescence. Although the small size of MSCs is known as a critical determinant of their fate, the main regulators of stem cell senescence and the underlying signaling have not been addressed. Umbilical cord blood-derived MSCs (UCB-MSCs) were obtained using size-isolation methods and then cultured with control or small cells to investigate the major factors that modulate MSC senescence. Cytokine array data suggested that the secretion of interukin-8 (IL-8) or growth-regulated oncogene-alpha (GROa) by senescent cells was markedly inhibited during incubation of small cells along with suppression of cognate receptor (C-X-C motif chemokine receptor2, CXCR2) via blockade of the autocrine/paracrine positive loop. Moreover, signaling via toll-like receptor 2 (TLR2) and TLR5, both pattern recognition receptors, drove cellular senescence of MSCs, but was inhibited in small cells. The activation of TLRs (2 and 5) through ligand treatment induced a senescent phenotype in small cells. Collectively, our data suggest that small cell from UCB-MSCs exhibit delayed cellular senescence by inhibiting the process of TLR signaling-mediated senescence-associated secretory phenotype (SASP) activation.

Long-term cultured mesenchymal stem cells frequently develop genomic mutations but do not undergo malignant transformation

Cytotherapy ◽  
2014 ◽  
Vol 16 (4) ◽  
pp. S75-S76
Author(s):  
Y. Wang ◽  
Z. Han ◽  
Z. Zhang ◽  
Y. Chi ◽  
Z. Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Malignant Transformation

Evaluation of Canine Bone Marrow-derived Mesenchymal Stem Cells After Long-term Cryopreservation

2013 ◽  
Vol 30 (12) ◽  
pp. 1032-1037 ◽  
Author(s):  
Xiaofeng Zhu ◽  
Fang Yuan ◽  
Houxuan Li ◽  
Yuqian Zheng ◽  
Yin Xiao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells

Enhancing therapeutic effects and in vivo tracking of adipose tissue derived mesenchymal stem cells for liver injury using bioorthogonal click chemistry

Nanoscale ◽  
2020 ◽  
Author(s):  
Naishun Liao ◽  
Da Zhang ◽  
Ming Wu ◽  
Huang-Hao Yang ◽  
Xiaolong Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Stem Cell ◽  
Liver Injury ◽  
Mesenchymal Stem Cell ◽  
Liver Diseases ◽  
Therapeutic Effects

Adipose tissue derived mesenchymal stem cell (ADSC)-based therapy is attractive for liver diseases, but the long-term therapeutic outcome is still far from satisfaction due to low hepatic engraftment efficiency of...

scholarly journals Influence of Egr-1 in Cardiac Tissue-Derived Mesenchymal Stem Cells in Response to Glucose Variations

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Daniela Bastianelli ◽  
Camilla Siciliano ◽  
Rosa Puca ◽  
Andrea Coccia ◽  
Colin Murdoch ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Target Genes ◽  
Cardiac Tissue ◽  
Early Response ◽  
Induced Response ◽  
Protein Levels ◽  
Short Period ◽  
Phenotype Analysis

Mesenchymal stem cells (MSCs) represent a promising cell population for cell therapy and regenerative medicine applications. However, how variations in glucose are perceived by MSC pool is still unclear. Since, glucose metabolism is cell type and tissue dependent, this must be considered when MSCs are derived from alternative sources such as the heart. The zinc finger transcription factor Egr-1 is an important early response gene, likely to play a key role in the glucose-induced response. Our aim was to investigate how short-term changes inin vitroglucose concentrations affect multipotent cardiac tissue-derived MSCs (cMSCs) in a mouse model of Egr-1 KO (Egr-1−/−). Results showed that loss of Egr-1 does not significantly influence cMSC proliferation. In contrast, responses to glucose variations were observed in wt but not in Egr-1−/−cMSCs by clonogenic assay. Phenotype analysis by RT-PCR showed that cMSCs Egr-1−/−lost the ability to regulate the glucose transporters GLUT-1 and GLUT-4 and, as expected, the Egr-1 target genes VEGF, TGFβ-1, and p300. Acetylated protein levels of H3 histone were impaired in Egr-1−/−compared to wt cMSCs. We propose that Egr-1 acts as immediate glucose biological sensor in cMSCs after a short period of stimuli, likely inducing epigenetic modifications.

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