scholarly journals Multipotent skin-derived precursors: adult neural crest-related precursors with therapeutic potential

2007 ◽  
Vol 363 (1489) ◽  
pp. 185-198 ◽  
Author(s):  
Karl J.L Fernandes ◽  
Jean G Toma ◽  
Freda D Miller
Keyword(s):  
Neural Crest ◽  
Therapeutic Potential ◽  
Precursor Cell ◽  
Cell Type ◽  
Differentiation Potential ◽  
Therapeutic Implications ◽  
Lineage Differentiation ◽  
Mammalian Skin ◽  
Surprising Finding

We previously made the surprising finding that cultures of multipotent precursors can be grown from the dermis of neonatal and adult mammalian skin. These skin-derived precursors (SKPs) display multi-lineage differentiation potential, producing both neural and mesodermal progeny in vitro , and are an apparently novel precursor cell type that is distinct from other known precursors within the skin. In this review, we begin by placing these findings within the context of the rapidly evolving stem cell field. We then describe our recent efforts focused on understanding the developmental biology of SKPs, discussing the idea that SKPs are neural crest-related precursors that (i) migrate into the skin during embryogenesis, (ii) persist within a specific dermal niche, and (iii) play a key role in the normal physiology, and potentially pathology, of the skin. We conclude by highlighting some of the therapeutic implications and unresolved questions raised by these studies.

Abstract P014: Oxidative Stress Mediated Regulation of Antioxidants in Cardiac Progenitor Cells

2011 ◽  
Vol 109 (suppl_1) ◽  
Author(s):  
Gokulakrishnan Iyer ◽  
Michael E Davis
Keyword(s):  
Oxidative Stress ◽  
Xanthine Oxidase ◽  
Progenitor Cells ◽  
Cardiac Progenitor Cells ◽  
Dependent Manner ◽  
Akt Inhibitor ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Phosphorylated Akt

Cardiac diseases are the leading causes of death throughout the world and transplantation of endogenous myocardial progenitor population with robust cardiovascular lineage differentiation potential is a promising therapeutic strategy. Therefore, in vitro expansion and transplantation of cardiac progenitor cells (CPCs) is currently in early clinical testing as a potential treatment for severe cardiac dysfunction. However, poor survival and engraftment of cells is one of the major limitations of cell transplantation therapy. Oxidative stress is increased in the ischemic myocardium and indirect inferences suggest the vulnerability of CPCs to oxidative stress. In this study, we show that in vitro, resident c-kit positive CPCs isolated from rat myocardium are significantly (p<0.05) resistant to superoxide-induced apoptosis compared to cardiomyocytes as analyzed by the number of sub-G1 population following xanthine/xanthine oxidase treatment. Interestingly, CPCs have two to four fold higher basal SOD1 and SOD2 activities (p<0.01) compared to cardiomyocytes and endothelial cells. Superoxide treatment increased expression of SOD1 (p<0.01), SOD2 (p<0.01), and glutathione peroxidase (p<0.05) mRNAs within 6 h of treatment compared to control cells. Recent studies suggest the involvement of AKT in controlling cell death, survival and also expression of SOD enzymes. Therefore, we investigated the involvement of AKT in CPCs subjected to oxidative stress. Western blot analysis revealed that the amount of phosphorylated AKT increased significantly within 10 minutes of xanthine/xanthine oxidase treatment. In addition, treatment with LY294002 - a PI3 kinase/AKT inhibitor, increased apoptosis in CPCs treated with superoxide. Our studies demonstrate a novel finding in which resident progenitor cells are protected from oxidative injury by containing higher basal levels of antioxidants as compared to myocytes. Moreover, under oxidant challenge antioxidant levels are regulated, possibly in an AKT-dependent manner. Further elucidation of this pathway may lead to novel therapeutic opportunities.

scholarly journals Soft Substrate Maintains Proliferative and Adipogenic Differentiation Potential of human Mesenchymal Stem Cells on Long Term Expansion by Delaying Senescence

2018 ◽  
Author(s):  
Sanjay K. Kureel ◽  
Pankaj Mogha ◽  
Akshada Khadpekar ◽  
Vardhman Kumar ◽  
Rohit Joshi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Culture System ◽  
Human Mesenchymal Stem Cells ◽  
Population Doubling ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Poly Acrylamide

AbstractHuman mesenchymal stem cells (hMSCs), when cultured on tissue culture plate (TCP) for in vitro expansion, they spontaneously lose their proliferative capacity and multi-lineage differentiation potential. They also lose their distinct spindle morphology and become large and flat. After a certain number of population doubling, they enter into permanent cell cycle arrest, called senescence. This is a major roadblock for clinical use of hMSCs which demands large number of cells. A cell culture system is needed which can maintain the stemness of hMSCs over long term passages yet simple to use. In this study, we explore the role of substrate rigidity in maintaining stemness. hMSCs were serially passaged on TCP and 5 kPa poly-acrylamide gel for 20 population doubling. It was found that while on TCP, cell growth reached a plateau at cumulative population doubling (CPD) = 12.5, on 5 kPa gel, they continue to proliferate linearly till we monitored (CPD = 20). We also found that while on TCP, late passage MSCs lost their adipogenic potential, the same was maintained on soft gel. Cell surface markers related to MSCs were also unaltered. We demonstrated that this maintenance of stemness was correlated with delay in onset of senescence, which was confirmed by β-gal assay and by differential expression of vimentin, Lamin A and Lamin B. As preparation of poly-acrylamide gel is a simple, well established, and well standardized protocol, we believe that this system of cell expansion will be useful in therapeutic and research applications of hMSCs.One Sentence SummaryhMSCs retain their stemness when expanded in vitro on soft polyacrylamide gel coated with collagen by delaying senescence.Significance StatementFor clinical applications, mesenchymal stem cells (MSCs) are required in large numbers. As MSCs are available only in scarcity in vivo, to fulfill the need, extensive in vitro expansion is unavoidable. However, on expansion, they lose their replicative and multi-lineage differentiation potential and become senescent. A culture system that can maintain MSC stemness on long-term expansion, without compromising the stemness, is need of the hour. In this paper, we identified polyacrylamide (PAA) hydrogel of optimum stiffness that can be used to maintain stemness of MSCs during in vitro long term culture. Large quantity of MSCs thus grown can be used in regenerative medicine, cell therapy, and in treatment of inflammatory diseases.

Proliferation and differentiation potential of rat forebrain oligodendroglial progenitors both in vitro and in vivo

Development ◽  
1991 ◽  
Vol 111 (4) ◽  
pp. 1061-1080 ◽  
Author(s):  
R. Hardy ◽  
R. Reynolds
Keyword(s):  
Progenitor Cell ◽  
Neonatal Rat ◽  
Myelin Proteins ◽  
Brdu Incorporation ◽  
Cell Type ◽  
Differentiation Potential ◽  
Serum Free ◽  
Rat Forebrain

We have followed the development of the O-2A progenitor cell from the neonatal rat forebrain, both in dissociated cell culture and in cryostat sections, using immunocytochemical techniques employing a panel of antibodies that recognise the cells at different stages of their development. This included the monoclonal antibody LB1, which binds to the surface ganglioside GD3 expressed on O-2A progenitor cells. In secondary cultures enriched for O-2A progenitors maintained in a serum-free chemically defined medium, a large proportion of the cells are primed to differentiate into oligodendroglia and go on to express the oligodendroglial specific surface glycolipid galactocerebroside (GC) and then the myelin proteins CNP and MBP. However, a significant proportion of immature bipolar GD3+ cells remained after 6 days in secondary culture. It appears that not all the O-2A progenitors in our cultures differentiate immediately and some cells remain in an undifferentiated state and divide to replenish progenitor numbers. We have also identified in our cultures a small apolar GD3- cell, which when isolated differentiated into a GD3+ bipolar O-2A progenitor cell. We have termed this cell type a preprogenitor. The differentiation of this cell type into O-2A progenitors may be the source of the immature GD3+ cells present at the later stages of our secondary cultures. The proliferative profile of the cultures was studied using 5′bromo-2-deoxyuridine (BrdU) incorporation as an index of mitosis. Only the immature, bipolar O-2A progenitors were seen to divide at any time in serum-free culture. Neither the more mature multipolar O-2A cells nor the oligodendroglia were seen to divide. The developmental profile of the O-2A cells in the rat forebrain in vivo showed a largely similar progression to that in culture, with a time lag of at least 6 days between GD3 expression and the onset of myelination. BrdU incorporation studies in vivo also showed that the GD3+ progenitor cell is mitotic whereas the GC(+)-expressing oligodendroglia is not. We have shown that there are several significant alterations in the timing of antigen expression in both O-2A progenitors and oligodendroglia in vitro compared to that seen in vivo.

scholarly journals ANTIOXIDANT, CYTOTOXICITY, AND STABILITY EVALUATION OF GINKGO BILOBA EXTRACT-BASED MICROEMULSIONS FOR ENHANCED THERAPEUTIC ACTIVITY

2017 ◽  
Vol 10 (8) ◽  
pp. 335 ◽  
Author(s):  
Manisha Singh ◽  
Surya Pratap Singh ◽  
Rachana R
Keyword(s):  
Ginkgo Biloba ◽  
Therapeutic Potential ◽  
Ginkgo Biloba Extract ◽  
In Vitro Cytotoxicity ◽  
Scavenging Activity ◽  
Therapeutic Implications ◽  
Chromatography Analysis ◽  
Ic50 Value ◽  
Cytotoxicity Assessment

Objective: This study is aimed to evaluate the antioxidant (AO) potential, cytotoxicity, and stability of preformulated Ginkgo biloba standard extractmicroemulsion (GBME), to investigate if, it retains the therapeutic potential of EGB761 and remains safe and stable for a longer period.Method and Results: GBME has shown enhanced AO (85.2±0.78%, IC50=31.3±0.45 μg/ml) in comparison to EGB761 (74.1±0.51%,IC50=49.4±0.05 μg/ml) using 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assay. Similarly, 2,2-diphenyl-1-picryl-hydrazyl-hydrate(DPPH) assay has also shown that AO for GBME (94.6±0.04%, IC50=11.4±1.03 μg/ml) was higher than EGB761 (78.6±1.20%, IC50=34.6±0.81 μg/ml).Further, IC50 value of antiradical unit of GBME was much lesser (ABTS=14.3±1.05 μg/ml and DPPH=17.03±1.8 μg/ml) in comparison to EGB761(ABTS=34.1±1.62 μg/ml and DPPH=37.5±0.08 μg/ml). Equivalently, both, hydrogen peroxide scavenging activity, and nitric oxide activity wereappreciably higher for GBME than the pure extract. The in vitro cytotoxicity assessment showed that GBME is quite safe (98.68±0.76% cell viability) incomparison to EGB761 (83.29±1.02%). Thereafter, these samples were tested for stability by evaluating their AO activity along with high-performanceliquid chromatography analysis, for the major phytocompounds, after 1 year, and results suggested that AO of GBME remained stable while comparingwith the freshly prepared GBME, whereas AO of EGB761 reduced significantly as compared to freshly taken EGB761 extract implying the degradationof phytocompounds supporting decrease in AO activity.Conclusion: Therefore, the observed results suggest that GBME maintained AO and scavenging activity along with enhanced shelf life with no observedtoxicity, which can be explored further for its potential therapeutic implications in various oxidative stress-induced central nervous system disorders.

scholarly journals Nuclear reprogramming in cell–free extracts

2003 ◽  
Vol 358 (1436) ◽  
pp. 1389-1395 ◽  
Author(s):  
Philippe Collas
Keyword(s):  
Somatic Cell ◽  
Adult Stem Cells ◽  
Regulatory Function ◽  
Nuclear Reprogramming ◽  
Cell Type ◽  
Differentiation Potential ◽  
Chromatin Remodelling Complex ◽  
Therapeutic Applications ◽  
Somatic Cell Type

Methods for directly turning a somatic cell type into another type (a process referred to as transdifferentiation) would be beneficial for producing replacement cells for therapeutic applications. Adult stem cells have been shown to display a broader differentiation potential than anticipated and may contribute to tissues other than those in which they reside. In addition, novel transdifferentiation strategies are being developed. I report recent results on the functional reprogramming of a somatic cell using a nuclear and cytoplasmic extract derived from another somatic cell type. The reprogramming of 293T fibroblasts in an extract from T cells is evidenced by nuclear uptake and the assembly of transcription factors, induction of activity of a chromatin remodelling complex, changes in chromatin composition and activation of lymphoid cell–specific genes. The reprogrammed cells express T–cell–specific surface molecules and a complex regulatory function. Reprogramming cells in cell–free extracts may create possibilities for producing replacement cells for therapeutic applications. The system may also constitute a powerful tool to examine the mechanisms of nuclear reprogramming, at least as they occur in vitro .

The roles of Frizzled-3 and Wnt3a on melanocyte development: In vitro studies on neural crest cells and melanocyte precursor cell lines

2014 ◽  
Vol 75 (2) ◽  
pp. 100-108 ◽  
Author(s):  
Chung-Hsing Chang ◽  
Rong-Kung Tsai ◽  
Ming-Hsien Tsai ◽  
Yi-Hsiung Lin ◽  
Tomohisa Hirobe
Keyword(s):  
Neural Crest ◽  
Cell Lines ◽  
Neural Crest Cells ◽  
Precursor Cell ◽  
In Vitro Studies ◽  
Development In Vitro

3D printable Sodium alginate-Matrigel (SA-MA) hydrogel facilitated ectomesenchymal stem cells (EMSCs) neuron differentiation

2020 ◽  
Vol 35 (6) ◽  
pp. 709-719 ◽  
Author(s):  
Yang Li ◽  
Xia Cao ◽  
Wenwen Deng ◽  
Qingtong Yu ◽  
Congyong Sun ◽  
...  
Keyword(s):  
Stem Cells ◽  
Sodium Alginate ◽  
Neuronal Differentiation ◽  
Adult Stem Cells ◽  
Three Dimensional ◽  
Cell Types ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Cord Injury

Ectomesenchymal stem cells (EMSCs) are typical adult stem cells obtained from the cranial neural crest. They have the potential to differentiate into various cell types, such as osseous cells, neurons and glial cells. Three-dimensional (3 D) printing is a novel method to construct biological structures by rapid prototyping. Previously, our group reported on the stemness and multi-lineage differentiation potential of EMSCs on gels. However, the exploration of EMSCs in 3 D printing and then evaluation of the growth and neuronal differentiation of EMSCs on extruded 3 D printable hybrid hydrogels has not been reported. Therefore, the current study explored the novel hybrid Sodium alginate-Matrigel (SA-MA) hydrogel extruded 3 D printing to design an in vitro scaffold to promote the differentiation and growth of EMSCs. In addition, the physical properties of the hydrogel were characterized and its drug-releasing property determined. Notably, the results showed that the construct exhibited a sustain-released effect of growth factor BDNF in accordance with the Higuchi equation. Moreover, the cell survival rate on the 3 D printed scaffold was 88.22 ± 1.13% with higher neuronal differentiation efficiency compared with 2 D culture. Thus, SA-MA’s ability to enhanced EMSCs neuronal differentiation offers a new biomaterial for neurons regeneration in the treatment of spinal cord injury.

scholarly journals An Efficient Protocol for Deriving Liver Stem Cells from Neonatal Mice: Validating Its Differentiation Potential

2015 ◽  
Vol 2015 ◽  
pp. 1-10
Author(s):  
Sugapriya Dhanasekaran ◽  
Devilakshmi Sithambaram ◽  
Kavitha Govarthanan ◽  
Bijesh Kumar Biswal ◽  
Rama S. Verma
Keyword(s):  
Stem Cells ◽  
Cell Types ◽  
Single Step ◽  
Stem Cell Markers ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Preclinical Trials ◽  
Neonatal Liver ◽  
Hepatocyte Markers

The success of liver regeneration depends on the availability of suitable cell types and their potential to differentiate into functional hepatocytes. To identify the stem cells which have the ability to differentiate into hepatocytes, we used neonatal liver as source. However, the current protocol for isolating stem cells from liver involves enzymes like collagenase, hyaluronidase exposed for longer duration which limits the success. This results in the keen interest to develop an easy single step enzyme digestion protocol for isolating stem cells from liver for tissue engineering approaches. Thus, the unlimited availability of cell type favors setting up the functional recovery of the damaged liver, ensuring ahead success towards treating liver diseases. We attempted to isolate liver stem derived cells (LDSCs) from mouse neonatal liver using single step minimal exposure to enzyme followed byin vitroculturing. The cells isolated were characterized for stem cell markers and subjected to lineage differentiation. Further, LDSCs were induced to hepatocyte differentiation and validated with hepatocyte markers. Finally, we developed a reproducible, efficient protocol for isolation of LDSCs with functional hepatocytes differentiation potential, which further can be used asin vitromodel system for assessing drug toxicity assays in various preclinical trials.

scholarly journals Cardiac Derived CD51-Positive Mesenchymal Stem Cells Enhance the Cardiac Repair Through SCF-Mediated Angiogenesis in Mice With Myocardial Infarction

2021 ◽  
Vol 9 ◽  
Author(s):  
Dong Mei Xie ◽  
Yang Chen ◽  
Yan Liao ◽  
Wanwen Lin ◽  
Gang Dai ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cells ◽  
Cardiac Fibrosis ◽  
Therapeutic Potential ◽  
Cardiac Repair ◽  
Surface Proteins ◽  
Differentiation Potential ◽  
Cardiac Wall

Objective: Many tissues contained resident mesenchymal stromal/stem cells (MSCs) that facilitated tissue hemostasis and repair. However, there is no typical marker to identify the resident cardiac MSCs. We aimed to determine if CD51 could be an optimal marker of cardiac MSCs and assess their therapeutic potential for mice with acute myocardial infarction (AMI).Methods: Cardiac-derived CD51+CD31–CD45–Ter119– cells (named CD51+cMSCs) were isolated from C57BL/6 mice(7-day-old) by flow cytometry. The CD51+cMSCs were characterized by proliferation capacity, multi-differentiation potential, and expression of typical MSC-related markers. Adult C57BL/6 mice (12-week-old) were utilized for an AMI model via permanently ligating the left anterior descending coronary artery. The therapeutic efficacy of CD51+cMSCs was estimated by echocardiography and pathological staining. To determine the underlying mechanism, lentiviruses were utilized to knock down gene (stem cell factor [SCF]) expression of CD51+cMSCs.Results: In this study, CD51 was expressed in the entire layers of the cardiac wall in mice, including endocardium, epicardium, and myocardium, and its expression was decreased with age. Importantly, the CD51+cMSCs possessed potent self-renewal potential and multi-lineage differentiation capacity in vitro and also expressed typical MSC-related surface proteins. Furthermore, CD51+cMSC transplantation significantly improved cardiac function and attenuated cardiac fibrosis through pro-angiogenesis activity after myocardial infarction in mice. Moreover, SCF secreted by CD51+cMSCs played an important role in angiogenesis both in vivo and in vitro.Conclusions: Collectively, CD51 is a novel marker of cardiac resident MSCs, and CD51+cMSC therapy enhances cardiac repair at least partly through SCF-mediated angiogenesis.

scholarly journals Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem CellsEx Vivo

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
June Seok Heo ◽  
Hyun Ok Kim ◽  
Seung Yong Song ◽  
Dae Hyun Lew ◽  
Youjeong Choi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cellular Therapy ◽  
Replicative Senescence ◽  
Therapeutic Potential ◽  
Global Gene Expression ◽  
S Phase ◽  
Global Gene Expression Profile ◽  
Rt Pcr ◽  
Differentiation Potential

Mesenchymal stem cells (MSCs) possess great therapeutic potential. Efficientin vitroexpansion of MSCs is however necessary for their clinical application. The extracellular matrix (ECM) provides structural and biochemical support to the surrounding cells, and it has been used as a coating substrate for cell culture. In this study, we have aimed to improve the functionality and stemness of MSCs during culture using poly-L-lysine (PLL). Functionality of MSCs was analysed by cell cycle analysis, differentiation assay,β-galactosidase staining, and RT-PCR. Furthermore, we assessed the global gene expression profile of MSCs on uncoated and PLL-coated plates. MSCs on PLL-coated plates exhibited a faster growth rate with increased S-phase and upregulated expression of the stemness markers. In addition, their osteogenic differentiation potential was increased, and genes involved in cell adhesion, FGF-2 signalling, cell cycle, stemness, cell differentiation, and cell proliferation were upregulated, compared to that of the MSCs cultured on uncoated plates. We also confirmed that MSCs on uncoated plates expressed higherβ-galactosidase than the MSCs on PLL-coated plates. We demonstrate that PLL provides favourable microenvironment for MSC culture by reversing the replicative senescence. This method will significantly contribute to effective preparation of MSCs for cellular therapy.

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