In VitroExpansion of Bone Marrow Derived Mesenchymal Stem Cells Alters DNA Double Strand Break Repair of Etoposide Induced DNA Damage

Mesenchymal stem cells (MSCs) are of interest for use in diverse cellular therapies.Ex vivoexpansion of MSCs intended for transplantation must result in generation of cells that maintain fidelity of critical functions. Previous investigations have identified genetic and phenotypic alterations of MSCs within vitropassage, but little is known regarding how culturing influences the ability of MSCs to repair double strand DNA breaks (DSBs), the most severe of DNA lesions. To investigate the response to DSB stress with passagein vitro, primary human MSCs were exposed to etoposide (VP16) at various passages with subsequent evaluation of cellular damage responses and DNA repair. Passage number did not affect susceptibility to VP16 or the incidence and repair kinetics of DSBs. Nonhomologous end joining (NHEJ) transcripts showed little alteration with VP16 exposure or passage; however, homologous recombination (HR) transcripts were reduced following VP16 exposure with this decrease amplified as MSCs were passagedin vitro. Functional evaluations of NHEJ and HR showed that MSCs were unable to activate NHEJ repair following VP16 stress in cells after successive passage. These results indicate thatex vivoexpansion of MSCs alters their ability to perform DSB repair, a necessary function for cells intended for transplantation.

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Current Strategies to Generate Human Mesenchymal Stem Cells In Vitro

Stem Cells International ◽

10.1155/2018/6726185 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 21

Author(s):

Jennifer Steens ◽

Diana Klein

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Current Knowledge ◽

Ex Vivo ◽

Human Mesenchymal Stem Cells ◽

Human Mscs ◽

Multipotent Stem Cells ◽

Adult Organism ◽

Almost All

Mesenchymal stem cells (MSCs) are heterogeneous multipotent stem cells that are involved in the development of mesenchyme-derived evolving structures and organs during ontogeny. In the adult organism, reservoirs of MSCs can be found in almost all tissues where MSCs contribute to the maintenance of organ integrity. The use of these different MSCs for cell-based therapies has been extensively studied over the past years, which highlights the use of MSCs as a promising option for the treatment of various diseases including autoimmune and cardiovascular disorders. However, the proportion of MSCs contained in primary isolates of adult tissue biopsies is rather low and, thus, vigorous ex vivo expansion is needed especially for therapies that may require extensive and repetitive cell substitution. Therefore, more easily and accessible sources of MSCs are needed. This review summarizes the current knowledge of the different strategies to generate human MSCs in vitro as an alternative method for their applications in regenerative therapy.

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DNA Methylation Changes duringIn VitroPropagation of Human Mesenchymal Stem Cells: Implications for Their Genomic Stability?

Stem Cells International ◽

10.1155/2013/192425 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 33

Author(s):

Angela Bentivegna ◽

Mariarosaria Miloso ◽

Gabriele Riva ◽

Dana Foudah ◽

Valentina Butta ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Ex Vivo ◽

Genomic Stability ◽

Great Promise ◽

Human Mscs ◽

Low Oxygen ◽

Long Term Culture ◽

Functional Changes

Mesenchymal stem cells (MSCs) hold great promise for the treatment of numerous diseases. A major problem for MSC therapeutic use is represented by the very low amount of MSCs which can be isolated from different tissues; thusex vivoexpansion is indispensable. Long-term culture, however, is associated with extensive morphological and functional changes of MSCs. In addition, the concern that they may accumulate stochastic mutations which lead the risk of malignant transformation still remains. Overall, the genome of human MSCs (hMSCs) appears to be apparently stable throughout culture, though transient clonal aneuploidies have been detected. Particular attention should be given to the use of low-oxygen environment in order to increase the proliferative capacity of hMSCs, since data on the effect of hypoxic culture conditions on genomic stability are few and contradictory. Furthermore, specific and reproducible epigenetic changes were acquired by hMSCs duringex vivoexpansion, which may be connected and trigger all the biological changes observed. In this review we address current issues on long-term culture of hMSCs with a 360-degree view, starting from the genomic profiles and back, looking for an epigenetic interpretation of their genetic stability.

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Exploring the Role of Mesenchymal Stem Cells During Normothermic Organ Perfusion: A New Paradigm to Enhance Outcome Following Allograft Transplantation

The Open Stem Cell Journal ◽

10.2174/1876893801805010047 ◽

2018 ◽

Vol 5 (1) ◽

pp. 47-52

Author(s):

Mohamed Morsy ◽

Mohammad Ayaz Hossain ◽

Atul Bagul

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Organ Transplantation ◽

Ex Vivo ◽

Solid Organ Transplantation ◽

Short Review ◽

Solid Organ ◽

New Paradigm ◽

Liver And Kidney

Background: Normothermic Machine Perfusion (NMP) has been established in the field of solid organ transplantation for both liver and kidney allografts. The ability to perfuse organs at body temperature enables viability assessment as well as optimisation prior to implantation. Discussion: A recent in vitro report of the use of Mesenchymal Stem Cells (MSCs) in the use of a normothermic lung perfusion circuit has raised the possibility of their use in solid organ transplantation. The aim of this short review is to outline the potential uses of bone marrow derived MSCs for their use in renal allograft ex vivo NMP. An overview is provided of current literature of NMP as well as theorised uses for MSCs.

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Transduction of Bone-Marrow-Derived Mesenchymal Stem Cells by Using Lentivirus Vectors Pseudotyped with Modified RD114 Envelope Glycoproteins

Journal of Virology ◽

10.1128/jvi.78.3.1219-1229.2004 ◽

2004 ◽

Vol 78 (3) ◽

pp. 1219-1229 ◽

Cited By ~ 96

Author(s):

Xian-Yang Zhang ◽

Vincent F. La Russa ◽

Jakob Reiser

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Ex Vivo ◽

Leukemia Virus ◽

Human Mscs ◽

Systemic Delivery ◽

Endogenous Virus ◽

Hiv 1

ABSTRACT Bone-marrow-derived mesenchymal stem cells (MSCs) have attracted considerable attention as tools for the systemic delivery of therapeutic proteins in vivo, and the ability to efficiently transfer genes of interest into such cells would create a number of therapeutic opportunities. We have designed and tested a series of human immunodeficiency virus type 1 (HIV-1)-based vectors and vectors based on the oncogenic murine stem cell virus to deliver and express transgenes in human MSCs. These vectors were pseudotyped with either the vesicular stomatitis virus G (VSV-G) glycoprotein (GP) or the feline endogenous virus RD114 envelope GP. Transduction efficiencies and transgene expression levels in MSCs were analyzed by quantitative flow cytometry and quantitative real-time PCR. While transduction efficiencies with virus particles pseudotyped with the VSV-G GP were found to be high, RD114 pseudotypes revealed transduction efficiencies that were 1 to 2 orders of magnitude below those observed with VSV-G pseudotypes. However, chimeric RD114 GPs, with the transmembrane and extracellular domains fused to the cytoplasmic domain derived from the amphotropic Moloney murine leukemia virus 4070A GP, revealed about 15-fold higher titers relative to the unmodified RD114 GP. The transduction efficiencies in human MSCs of HIV-1-based vectors pseudotyped with the chimeric RD114 GP were similar to those obtained with HIV-1 vectors pseudotyped with the VSV-G GP. Our results also indicate that RD114 pseudotypes were less toxic than VSV-G pseudotypes in human MSC progenitor assays. Taken together, these results suggest that lentivirus pseudotypes bearing alternative Env GPs provide efficient tools for ex vivo modification of human MSCs.

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Thermoreversible hyaluronan-based hydrogel supports in vitro and ex vivo disc-like differentiation of human mesenchymal stem cells

The Spine Journal ◽

10.1016/j.spinee.2013.05.029 ◽

2013 ◽

Vol 13 (11) ◽

pp. 1627-1639 ◽

Cited By ~ 52

Author(s):

Marianna Peroglio ◽

David Eglin ◽

Lorin M. Benneker ◽

Mauro Alini ◽

Sibylle Grad

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Ex Vivo ◽

Human Mesenchymal Stem Cells

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Human mesenchymal stem cells inhibit differentiation and function of monocyte-derived dendritic cells

Blood ◽

10.1182/blood-2004-02-0586 ◽

2005 ◽

Vol 105 (10) ◽

pp. 4120-4126 ◽

Cited By ~ 861

Author(s):

Xiao-Xia Jiang ◽

Yi Zhang ◽

Bing Liu ◽

Shuang-Xi Zhang ◽

Ying Wu ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Dendritic Cells ◽

Human Mesenchymal Stem Cells ◽

Interleukin 4 ◽

Human Mscs ◽

Gm Csf ◽

Macrophage Colony ◽

And Function

AbstractMesenchymal stem cells (MSCs), in addition to their multilineage differentiation, have a direct immunosuppressive effect on T-cell proliferation in vitro. However, it is unclear whether they also modulate the immune system by acting on the very first step. In this investigation, we addressed the effects of human MSCs on the differentiation, maturation, and function of dendritic cells (DCs) derived from CD14+ monocytes in vitro. Upon induction with granulocyte-macrophage colony-stimulating factor (GM-CSF) plus interleukin-4 (IL-4), MSC coculture could strongly inhibit the initial differentiation of monocytes to DCs, but this effect is reversible. In particular, such suppression could be recapitulated with no intercellular contact at a higher MSC/monocyte ratio (1:10). Furthermore, mature DCs treated with MSCs were significantly reduced in the expression of CD83, suggesting their skew to immature status. Meanwhile, decreased expression of presentation molecules (HLA-DR and CD1a) and costimulatory molecules (CD80 and CD86) and down-regulated IL-12 secretion were also observed. In consistence, the allostimulatory ability of MSC-treated mature DCs on allogeneic T cells was impaired. In conclusion, our data suggested for the first time that human MSCs could suppress monocyte differentiation into DCs, the most potent antigen-presenting cells (APCs), thus indicating the versatile regulation of MSCs on the ultimate specific immune response.

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Mesenchymal stem cells are injured by complement after their contact with serum

Blood ◽

10.1182/blood-2012-03-420612 ◽

2012 ◽

Vol 120 (17) ◽

pp. 3436-3443 ◽

Cited By ~ 94

Author(s):

Yan Li ◽

Feng Lin

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Surface ◽

Complement Activation ◽

Human Mscs ◽

Cellular Injury ◽

Allogeneic Mscs ◽

Complement Regulators

Abstract Despite the potent immunosuppressive activity that mesenchymal stem cells (MSCs) display in vitro, recent clinical trial results are disappointing, suggesting that MSC viability and/or function are greatly reduced after infusion. In this report, we demonstrated that human MSCs activated complement of the innate immunity after their contact with serum. Although all 3 known intrinsic cell-surface complement regulators were present on MSCs, activated complement overwhelmed the protection of these regulators and resulted in MSCs cytotoxicity and dysfunction. In addition, autologous MSCs suffered less cellular injury than allogeneic MSCs after contacting serum. All 3 complement activation pathways were involved in generating the membrane attack complex to directly injure MSCs. Supplementing an exogenous complement inhibitor, or up-regulating MSC expression levels of CD55, one of the cell-surface complement regulators, helped to reduce the serum-induced MSC cytotoxicity. Finally, adoptively transferred MSCs in complement deficient mice or complement-depleted mice showed reduced cellular injury in vivo compared with those in wild type mice. These results indicate that complement is integrally involved in recognizing and injuring MSCs after their infusion, suggesting that autologous MSCs may have ad-vantages over allogeneic MSCs, and that inhibiting complement activation could be a novel strategy to improve existing MSC-based therapies.

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Distribution of Seeded Mesenchymal Stem Cells on Hydroxyapatite Porous Ceramics

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.330-332.1141 ◽

2007 ◽

Vol 330-332 ◽

pp. 1141-1144 ◽

Cited By ~ 1

Author(s):

Mika Tadokoro ◽

Noriko Kotobuki ◽

Akira Oshima ◽

Hajime Ohgushi

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

In Vitro Culture ◽

Porous Ceramic ◽

Porous Ceramics ◽

Human Mscs ◽

Ceramic Scaffold ◽

Before And After

This study focused on in vivo osteogenic capability of bone marrow mesenchymal stem cells (MSCs) seeded on ceramic scaffold. Human MSCs from a single donor were seeded on hydroxyapatite porous ceramic (HAP) and were induced to the osteogenic lineage during in vitro culture condition, then the MSCs/HAP composites were implanted subcutaneously into immunodeficient rats. The cellular activities of the composites were assayed in order to evaluate the distribution and differentiation capability of seeded MSCs before and after implantation. These results showed that the new bone, after implantation, was derived from the donor MSCs, which adhered to the surface of the ceramics pore areas during in vitro culture. Therefore, the engrafted donor cells proliferated and showed continuous osteogenic differentiation within the recipients. Consequently, our study demonstrates the usefulness of MSCs/HAP composites for clinical applications.

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Aberrant PARP1 Activity Couples DNA Breaks to Deregulated Presynaptic Calcium Signalling and Lethal Seizures

10.1101/431916 ◽

2018 ◽

Cited By ~ 1

Author(s):

Emilia Komulainen ◽

Jack Badman ◽

Stephanie Rey ◽

Stuart Rulten ◽

Limei Ju ◽

...

Keyword(s):

Hippocampal Neurons ◽

Ex Vivo ◽

Calcium Signalling ◽

Strand Break ◽

Parp Inhibition ◽

Dna Breaks ◽

Single Strand Break ◽

Presynaptic Calcium ◽

Dna Strand

AbstractDefects in DNA single-strand break repair result in cerebellar ataxia which in Xrcc1Nes-Cre mice is promoted by hyperactivity of the DNA strand break sensor protein, Parp1. Here, we show that Parp1 hyperactivity extends beyond the cerebellum in Xrcc1-defective brain, resulting in lethal seizures and shortened lifespan. We demonstrate that aberrant Parp1 activation triggers seizure-like activity in Xrcc1-defective hippocampus ex vivo and aberrant presynaptic calcium signalling in isolated hippocampal neurons in vitro. Moreover, we show that these defects are prevented by Parp1 inhibition and/or deletion. Collectively, these data identify aberrant Parp1 activity at unrepaired DNA breaks as a cell-autonomous source of deregulated presynaptic calcium signalling, and highlight PARP inhibition as a possible therapeutic approach in XRCC1-mutated neurodegenerative disease.SummaryPARP1 activity and presynaptic Ca2+ signalling

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Rheb1-Deficient Neutrophils Promote Hematopoietic Stem/Progenitor Cell Proliferation via Mesenchymal Stem Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.650599 ◽

2021 ◽

Vol 9 ◽

Author(s):

Juan Gao ◽

Shuaibing Hou ◽

Shengnan Yuan ◽

Yuxia Wang ◽

Yanan Gao ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Myeloid Cells ◽

Human Mscs ◽

Hematopoietic Stem ◽

Progenitor Cell Proliferation ◽

High Level ◽

Lower Expression

Myeloid cells have been identified as hematopoietic stem cell (HSC)-regulating cells. However, the mechanisms by which myeloid cells regulate the function of HSCs are not fully defined. Our previous study indicated that the HSCs are over-expanded in Vav1-Cre;Rheb1fl/fl mice. Here, using in vivo and in vitro models, we found that Rheb1-deficient neutrophils remodeled the bone marrow environment and induced expansion of HSCs in vivo. Further studies showed that loss of Rheb1 impaired neutrophils’ ability to secrete IL-6, led mesenchymal stem cells (MSCs) to produce more SCF, and promote HSC proliferation. We further found that IL-6 suppressed SCF mRNA expression in human MSCs. Interesting, the high level of IL-6 was also related with poor survival of chronic myeloid leukemia (CML) patients, and higher expression of IL-6 in CML cells is associated with the lower expression of SCF in MSCs in patients. Our studies suggested that blocking IL-6 signaling pathway might stimulate MSCs to secrete more SCF, and to support hematopoietic stem/progenitor cells proliferation.

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