Behavior of mesenchymal stem cells stained with 4’, 6-diamidino-2-phenylindole dihydrochloride (DAPI) in osteogenic and non osteogenic cultures

Ocarino, Natália de Melo; Bozzi, Adriana; Pereira, Rafael Diogo; Breyner, Natália Martins; Silva, V L; Castanheira, Paula; Góes, Alfredo M.; Serákides, Rogéria

doi:10.32604/biocell.2008.32.175

Cited by 14 publications

(4 citation statements)

References 28 publications

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“…In brief, sterile DAPI stock solution (Sigma-Aldrich, USA) was added to the culture medium at the day of transplantation at a final concentration of 50 μg/ml for 2 h. After incorporation of the DAPI into the nuclei, cells were rinsed six times in PBS to remove unbounded DAPI, detached with 0.25% trypsin-EDTA, and resuspended in saline for cell transplantation. Using this protocol, the level of expression of DAPI in the MSCs is approximately 100% (25).…”

Section: Methodsmentioning

confidence: 99%

Time-Dependent Migration of Systemically Delivered Bone Marrow Mesenchymal Stem Cells to the Infarcted Heart

et al. 2010

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In this study the time course of homing and the body distribution of systemically delivered bone marrow mesenchymal stem cells (BM-MSCs) after myocardial infarction (MI) were evaluated. BM-MSCs were isolated from Wistar rats, expanded in vitro, and their phenotypical characterization was performed by flow cytometer. Rats were randomly divided into three groups: control, sham MI, and MI. BM-MSCs (5 × 10 6 ) were labeled with 99m Tc-HMPAO and injected through the tail vein 7 days after MI. Gamma camera imaging was performed at 5, 15, 30, and 60 min after cell inoculation. Due to the 99m Tc short half-life, cell migration and location were also evaluated in heart sections using DAPI-labeled cells 7 days after transplantation. Phenotypical characterization showed that BM-MSCs were CD90 + , CD73 + , CD54+ , and CD45 − . Five minutes after 99m Tc-HMPAO-labeled cell injection, they were detected in various tissues. The cells migrated mainly to the lungs (approximately 70%) and, in small amounts, to the heart, kidneys, spleen, and bladder. The number of cells in the heart and lungs decreased after 60 min. MI markedly increased the amount of cells in the heart, but not in the lungs, during the period of observation (4.55 ± 0.32 vs. 6.34 ± 0.67% of uptake in infarcted hearts). No significant differences were observed between control and sham groups. Additionally, 7 days after DAPI-labeled cells injection, they were still detected in the heart but only in infarcted areas. These results suggest that the migration of systemically delivered BM-MSCs to the heart is time dependent and MI specifically increases BM-MSCs homing to injured hearts. However, the systemic delivery is limited by cell entrapment in the lungs.Key words: Myocardial infarction; Mesenchymal stem cells; Homing; Body distribution INTRODUCTIONstem cells (BM-MSCs) that can differentiate into different cellular types (5,14,27). Many reports have shown that cellular therapy using Mesenchymal stem cells (MSCs) have been isolated from multiple adult tissue sources, such as cord blood, BM-MSCs is able to improve cardiac function after myocardial ischemia in different species, including huplacenta, adipose and dermal tissues, synovial fluid, deciduous teeth, and amniotic fluid (19). This broad distrimans (2,30). However, the mechanisms by which BMMSCs induce their beneficial effects in the heart are still bution of sources combined with their ability to differentiate into multiple mesenchymal phenotypes, such as controversial. Thus, it has been proposed that BM-MSCs can differentiate into both vascular endothelial cells and bone, cartilage, tendon, and adipose tissue, has led them to be referred as potential therapeutic candidates for sevcardiomyocytes, activate local factors, fuse with resident cells, or even a combination of these mechanisms that eral diseases and degenerative processes, including myocardial infarction (MI) (18,37). In addition to these ultimately lead to a restoration of the cardiac structure and function (6,7,32 based therapy for myocardial repai...

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Section: Methodsmentioning

confidence: 99%

Time-Dependent Migration of Systemically Delivered Bone Marrow Mesenchymal Stem Cells to the Infarcted Heart

et al. 2010

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“…The use of the nontoxic dye PKH26 as a marker for identifying transplanted cells in vivo is well-established ( Jin et al, 2011 ; Liang et al, 2013 ). The fluorescent DNA dye DAPI has been used extensively for visualizing cell nuclei and is commonly used in immunocytochemistry applications, without changing cell viability and proliferation of MSCs ( Ocarino et al, 2008 ). In the present study, large numbers of PKH26/DAPI double-positive cells were observed in the injured hemisphere, which indicates that transplanted MSCs were able to survive in vivo for at least 3 days.…”

Section: Discussionmentioning

confidence: 99%

Therapeutic Benefits of Mesenchymal Stromal Cells in a Rat Model of Hemoglobin-Induced Hypertensive Intracerebral Hemorrhage

Ding¹,

Lin²,

Wei³

et al. 2017

Molecules and Cells

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Previous studies have shown that bone marrow mesenchymal stromal cell (MSC) transplantation significantly improves the recovery of neurological function in a rat model of intracerebral hemorrhage. Potential repair mechanisms involve anti-inflammation, anti-apoptosis and angiogenesis. However, few studies have focused on the effects of MSCs on inducible nitric oxide synthase (iNOS) expression and subsequent peroxynitrite formation after hypertensive intracerebral hemorrhage (HICH). In this study, MSCs were transplanted intracerebrally into rats 6 hours after HICH. The modified neurological severity score and the modified limb placing test were used to measure behavioral outcomes. Blood–brain barrier disruption and neuronal loss were measured by zonula occludens-1 (ZO-1) and neuronal nucleus (NeuN) expression, respectively. Concomitant edema formation was evaluated by H&E staining and brain water content. The effect of MSCs treatment on neuroinflammation was analyzed by immunohistochemical analysis or polymerase chain reaction of CD68, Iba1, iNOS expression and subsequent peroxynitrite formation, and by an enzyme-linked immunosorbent assay of pro-inflammatory factors (IL-1β and TNF-α). The MSCs-treated HICH group showed better performance on behavioral scores and lower brain water content compared to controls. Moreover, the MSC injection increased NeuN and ZO-1 expression measured by immunochemistry/immunofluorescence. Furthermore, MSCs reduced not only levels of CD68, Iba1 and pro-inflammatory factors, but it also inhibited iNOS expression and peroxynitrite formation in perihematomal regions. The results suggest that intracerebral administration of MSCs accelerates neurological function recovery in HICH rats. This may result from the ability of MSCs to suppress inflammation, at least in part, by inhibiting iNOS expression and subsequent peroxynitrite formation.

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“…Hematopoietic stem cells (HSCs) are capable of self-renewal and have the pluripotent ability to differentiate into all hematopoietic cell types (Yahara et al, 2020). DAPI (4′, 6-diamidino-2-phenylindole dihydrochloride) can be used to trace short periods of osteogenic differentiation of mesenchymal stem cells (Ocarino et al, 2008). HSCs give rise to oligopotent progenitor cells, such as common myeloid progenitor cells (CMPs), upon differentiation, which in turn, differentiate into macrophages and osteoclasts (Ono and Nakashima, 2018).…”

Section: Osteoblast Cell Differentiationmentioning

confidence: 99%

Effects of areca nut consumption on cell differentiation of osteoblasts, myoblasts, and fibroblasts

Chang¹

2023

Biocell

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Areca nut is used worldwide as a hallucinogenic addicting drug along the tropical belt. Arecoline, a toxic compound, is the most important alkaloid in areca nuts. The adverse effects of oral uptake and chewing of areca nut are well known. For example, the possibility of cancer caused by chewing areca nuts is widely discussed. Chewing areca nut has other adverse effects on other organs, including abnormal cell differentiation, oral cancer, and several other diseases. The use of areca nut is also associated with low birthweight. Skeletal musculature is the largest organ in the body and is attached to the bones. During embryo development, the differentiation of bone and muscle cells is critical. In this article, we reviewed the effects of areca nut and arecoline on embryonic cell differentiation, particularly osteoblasts, myoblasts, and fibroblasts.

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Behavior of mesenchymal stem cells stained with 4’, 6-diamidino-2-phenylindole dihydrochloride (DAPI) in osteogenic and non osteogenic cultures

Cited by 14 publications

References 28 publications

Time-Dependent Migration of Systemically Delivered Bone Marrow Mesenchymal Stem Cells to the Infarcted Heart

Time-Dependent Migration of Systemically Delivered Bone Marrow Mesenchymal Stem Cells to the Infarcted Heart

Therapeutic Benefits of Mesenchymal Stromal Cells in a Rat Model of Hemoglobin-Induced Hypertensive Intracerebral Hemorrhage

Effects of areca nut consumption on cell differentiation of osteoblasts, myoblasts, and fibroblasts

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