Showing 37 - 48 of 51 results for "05401"
11 Products
- ReferencePozzi S et al. (JUL 2006) Experimental hematology 34 7 934--42
Donor multipotent mesenchymal stromal cells may engraft in pediatric patients given either cord blood or bone marrow transplantation.
OBJECTIVE: Multipotent mesenchymal stromal cells (MSCs) are endowed with multilineage differentiative potential and immunomodulatory properties. It is still a matter of debate whether donor MSCs have sustained engraftment potential in host bone marrow (BM) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The aim of this study was to analyze the donor/recipient origin of MSCs in children receiving allogeneic either BM or cord blood (CB) transplantation. METHODS: Thirty-seven pediatric patients undergoing allo-HSCT for either a malignant or a nonmalignant disorder were enrolled in the study; 19 received CB and 18 BM transplantation. Results were compared with those obtained in 14 adults given BM transplantation for either malignant or nonmalignant disorders. MSCs were grown from BM aspirates obtained 1-17 and 2-192 months after allo-HSCT in pediatric and adult patients, respectively. MSC samples at the third-fourth passage were phenotypically characterized. Donor/recipient origin of MSCs was assessed by amelogenin assay and microsatellite analysis. RESULTS: MSCs could be grown from 30 of 37 children; at the third-fourth passage MSCs resulted positive (textgreater or = 98%) for CD73, CD105, CD106, CD29, CD13, CD44 and negative (textless or = 1%) for CD34, CD45, CD14. Mixed chimerism with donor cells was observed in 4 BM and 5 CB transplantation recipients, respectively; full recipient chimerism was detected in the remaining children. Full recipient MSC chimerism was observed also in all assessable (12/14) adult patients. CONCLUSIONS: BM of pediatric patients might be a more favorable milieu than that of adults for sustained engraftment of transplanted MSCs. MSCs able to engraft in the host can be transferred with cryopreserved CB units. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceSpaggiari GM et al. (FEB 2006) Blood 107 4 1484--90
Mesenchymal stem cell-natural killer cell interactions: evidence that activated NK cells are capable of killing MSCs, whereas MSCs can inhibit IL-2-induced NK-cell proliferation.
In recent years, mesenchymal stem cells (MSCs) have been shown to inhibit T-lymphocyte proliferation induced by alloantigens or mitogens. However, no substantial information is available regarding their effect on natural killer (NK) cells. Here we show that MSCs sharply inhibit IL-2-induced proliferation of resting NK cells, whereas they only partially affect the proliferation of activated NK cells. In addition, we show that IL-2-activated NK cells (but not freshly isolated NK cells) efficiently lyse autologous and allogeneic MSCs. The activating NK receptors NKp30, NKG2D, and DNAM-1 represented the major receptors responsible for the induction of NK-mediated cytotoxicity against MSCs. Accordingly, MSCs expressed the known ligands for these activating NK receptors-ULBPs, PVR, and Nectin-2. Moreover, NK-mediated lysis was inhibited when IFN-gamma-exposed MSCs were used as target cells as a consequence of the up-regulation of HLA class I molecules at the MSC surface. The interaction between NK cells and MSCs resulted not only in the lysis of MSCs but also in cytokine production by NK cells. These results should be taken into account when evaluating the possible use of MSCs in novel therapeutic strategies designed to improve engraftment or to suppress graft-versus-host disease (GVHD) in bone marrow transplantation. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceCorcione A et al. (JAN 2006) Blood 107 1 367--72
Human mesenchymal stem cells modulate B-cell functions.
Human mesenchymal stem cells (hMSCs) suppress T-cell and dendritic-cell function and represent a promising strategy for cell therapy of autoimmune diseases. Nevertheless, no information is currently available on the effects of hMSCs on B cells, which may have a large impact on the clinical use of these cells. hMSCs isolated from the bone marrow and B cells purified from the peripheral blood of healthy donors were cocultured with different B-cell tropic stimuli. B-cell proliferation was inhibited by hMSCs through an arrest in the G0/G1 phase of the cell cycle and not through the induction of apoptosis. A major mechanism of B-cell suppression was hMSC production of soluble factors, as indicated by transwell experiments. hMSCs inhibited B-cell differentiation because IgM, IgG, and IgA production was significantly impaired. CXCR4, CXCR5, and CCR7 B-cell expression, as well as chemotaxis to CXCL12, the CXCR4 ligand, and CXCL13, the CXCR5 ligand, were significantly down-regulated by hMSCs, suggesting that these cells affect chemotactic properties of B cells. B-cell costimulatory molecule expression and cytokine production were unaffected by hMSCs. These results further support the potential therapeutic use of hMSCs in immune-mediated disorders, including those in which B cells play a major role. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceBacigalupo A et al. (JUL 2005) Experimental hematology 33 7 819--27
T-cell suppression mediated by mesenchymal stem cells is deficient in patients with severe aplastic anemia.
OBJECTIVE: To compare the suppressive effect of mesenchymal stem cells (MSC), derived from normal individuals or severe aplastic anemia patients (SAA), on T-cell activation. PATIENTS AND METHODS: We studied bone marrow MSC from 19 healthy donors and 23 SAA patients in different phases of the disease: at diagnosis (n = 3), following immunosuppressive therapy (IS) (n = 16), or after a bone marrow transplant (BMT) (n = 4). MSC were tested for T-cell suppression in the following assays: mixed lymphocyte reaction (MLR), phytohemaglutinin (PHA)-primed cultures, activation surface markers, gamma-IFN production, hematopoietic colony formation (CFC), production of cyclic ADP-ribose (cADPR). RESULTS: The abnormalities of SAA MSC included: 1) significantly lower suppression of T-cell proliferation induced by alloantigens (p = 0.009) or PHA (p = 0.006); 2) impaired capacity to suppress CD38 expression on PHA-primed T cells (p = 0.001); 3) impaired ability to suppress gamma-IFN production in PHA cultures, resulting in an 11-fold higher gamma-IFN concentration; 4) no preventive effect on T cell-mediated inhibition of CFC; and 5) significantly reduced (p = 0.009) production of cADPR, a universal calcium mobilizer. MSC-mediated suppression of PHA-induced T-cell proliferation was restored to control levels in 3 of 4 patients post-BMT. CONCLUSION: The ability of MSC to downregulate T-cell priming, proliferation, and cytokine release is deficient in patients with SAA, persists indefinitely after immunosuppressive therapy, but seems to be restored after BMT. Whether these abnormalities are relevant to the pathogenesis of aplastic anemia remains to be determined. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceLee J-H et al. (JUL 2005) Experimental cell research 307 1 174--82
Contribution of human bone marrow stem cells to individual skeletal myotubes followed by myogenic gene activation.
Much attention is focused on characterizing the contribution of bone marrow (BM)-derived cells to regenerating skeletal muscle, fuelled by hopes for stem cell-mediated therapy of muscle degenerative diseases. Though physical integration of BM stem cells has been well documented, little evidence of functional commitment to myotube phenotype has been reported. This is due to the innate difficulty in distinguishing gene products derived from donor versus host nuclei. Here, we demonstrate that BM-derived stem cells contribute via gene expression following incorporation to skeletal myotubes. By co-culturing human BM-derived mesenchymal stem cells (MSC) with mouse skeletal myoblasts, physical incorporation was observed by genetic lineage tracing and species-specific immunofluorescence. We used a human-specific antibody against the intermediate filament protein nestin, a marker of regenerating skeletal muscle, to identify functional contribution of MSC to myotube formation. Although nestin expression was never detected in MSC, human-specific expression was detected in myotubes that also contained MSC-derived nuclei. This induction of gene expression following myotube integration suggests that bone marrow-derived stem cells can reprogram and functionally contribute to the muscle cell phenotype. We propose that this model of myogenic commitment may provide the means to further characterize functional reprogramming of MSC to skeletal muscle. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 15128 RosetteSep™ Human Mesenchymal Stem Cell Enrichment Cocktail Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 15128 Product Name: RosetteSep™ Human Mesenchymal Stem Cell Enrichment Cocktail - ReferenceSteward CG et al. (FEB 2005) Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation 11 2 115--21
High peripheral blood progenitor cell counts enable autologous backup before stem cell transplantation for malignant infantile osteopetrosis.
Autosomal recessive osteopetrosis (OP) is a rare, lethal disorder in which osteoclasts are absent or nonfunctional, resulting in a bone marrow cavity insufficient to support hematopoiesis. Because osteoclasts are derived from hematopoietic precursors, allogeneic hematopoietic cell transplantation can cure the bony manifestations of the disorder. However, high rates of graft failure have been observed in this population. It is not possible to harvest bone marrow from these patients for reinfusion should graft failure be observed. We report that 8 of 10 patients with OP had high numbers of circulating CD34(+) cells (3% +/- 0.9%). This increased proportion of peripheral CD34(+) cells made it possible to harvest 2 x 10(6) CD34(+) cells per kilogram with a total volume of blood ranging from 8.3 to 83.7 mL (1.3-11.6 mL/kg). In addition, colony-forming assays documented significantly more colony-forming unit-granulocyte-macrophage and burst-forming unit-erythroid in the blood of osteopetrotic patients compared with controls; the numbers of colony-forming units approximated those found in control marrow. We conclude that OP patients with high levels of circulating CD34(+) are candidates for peripheral blood autologous harvest by limited exchange transfusion. These cells are then available for reinfusion should graft failure be observed in patients for whom retransplantation is impractical. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceTauchmanovà et al. (FEB 2005) The Journal of clinical endocrinology and metabolism 90 2 627--34
Short-term zoledronic acid treatment increases bone mineral density and marrow clonogenic fibroblast progenitors after allogeneic stem cell transplantation.
Although osteoporosis is a relatively common complication after allogeneic stem cell transplantation, the role of bisphosphonates in its management has not yet been completely established. Thirty-two patients who underwent allogeneic stem cell transplantation were prospectively evaluated for bone mineral density (BMD) at the lumbar spine (LS) and femoral neck (FN) after a median period of 12.2 months. Then, 15 of the patients with osteoporosis or rapidly progressing osteopenia (bone loss textgreater 5%/yr) received three monthly doses of 4 mg zoledronic acid iv. Fifteen patients were followed up without treatment, and all 30 patients were reevaluated after 12 months for BMD and bone turnover markers. By using enriched mesenchymal stem cells in the colony-forming units fibroblast (CFU-F) assay, we evaluated the osteogenic stromal lineage. This procedure was performed in both groups of patients at study entry and after 12 months. The average BMD loss was 3.42% at LS and 3.8% at FN during a 1-yr longitudinal evaluation in 32 patients. Subsequently, BMD increased at both LS and FN (9.8 and 6.4%, respectively) in the zoledronic acid-treated cohort. Hydroxyproline excretion decreased, and serum bone-specific alkaline phosphatase increased significantly, whereas serum osteocalcin increase did not reach the limit of significance. A significant increase in CFU-F growth in vitro was induced by in vivo zoledronic acid administration. In the untreated group, no significant change was observed in bone turnover markers, LS BMD (-2.1%), FN BMD (-2.3%), and CFU-F colony number. In conclusion, short-term zoledronic acid treatment consistently improved both LS and FN BMD in transplanted patients who were at high risk for fast and/or persistent bone loss, partly by increasing the osteogenic progenitors in the stromal cell compartment. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceAhrens N et al. (SEP 2004) Transplantation 78 6 925--9
Mesenchymal stem cell content of human vertebral bone marrow.
Mesenchymal stem cells (MSCs) are capable of down-regulating alloimmune responses and promoting the engraftment of hematopoietic stem cells. MSCs may therefore be suitable for improving donor-specific tolerance induction in solid-organ transplantation. Cells from cadaveric vertebral bone marrow (V-BM), aspirated iliac crest-BM, and peripheral blood progenitor cells were compared. Cells were characterized by flow cytometry and colony assays. MSCs generated from V-BM were assayed for differentiation capacity and immunomodulatory function. A median 5.7 x 10(8) nucleated cells (NCs) were recovered per vertebral body. The mesenchymal progenitor, colony-forming unit-fibroblast, frequency in V-BM (11.6/10(5) NC, range: 6.0-20.0) was considerably higher than in iliac crest-BM (1.4/10(5) NC, range: 0.4-2.6) and peripheral blood progenitor cells (not detectable). MSC generated from V-BM had the typical MSC phenotype (CD105(pos)CD73(pos)CD45(neg)CD34(neg)), displayed multilineage differentiation potential, and suppressed alloreactivity in mixed lymphocyte reactions. V-BM may be an excellent source for MSC cotransplantation approaches. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceBieback K et al. (JAN 2004) Stem cells (Dayton, Ohio) 22 4 625--34
Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood.
Evidence has emerged that mesenchymal stem cells (MSCs) represent a promising population for supporting new clinical concepts in cellular therapy. However, attempts to isolate MSCs from umbilical cord blood (UCB) of full-term deliveries have previously either failed or been characterized by a low yield. We investigated whether cells with MSC characteristics and multi-lineage differentiation potential can be cultivated from UCB of healthy newborns and whether yields might be maximized by optimal culture conditions or by defining UCB quality criteria. Using optimized isolation and culture conditions, in up to 63% of 59 low-volume UCB units, cells showing a characteristic mesenchymal morphology and immune phenotype (MSC-like cells) were isolated. These were similar to control MSCs from adult bone marrow (BM). The frequency of MSC-like cells ranged from 0 to 2.3 clones per 1 x 10(8) mononuclear cells (MNCs). The cell clones proliferated extensively with at least 20 population doublings within eight passages. In addition, osteogenic and chondrogenic differentiation demonstrated a multi-lineage capacity comparable with BM MSCs. However, in contrast to MSCs, MSC-like cells showed a reduced sensitivity to undergo adipogenic differentiation. Crucial points to isolate MSC-like cells from UCB were a time from collection to isolation of less than 15 hours, a net volume of more than 33 ml, and an MNC count of more than 1 x 10(8) MNCs. Because MSC-like cells can be isolated at high efficacy from full-term UCB donations, we regard UCB as an additional stem cell source for experimental and potentially clinical purposes. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceOswald J et al. (JAN 2004) Stem cells (Dayton, Ohio) 22 3 377--84
Mesenchymal stem cells can be differentiated into endothelial cells in vitro.
Human bone marrow-derived mesenchymal stem cells (MSCs) have the potential to differentiate into mesenchymal tissues like osteocytes, chondrocytes, and adipocytes in vivo and in vitro. The aim of this study was to investigate the in vitro differentiation of MSCs into cells of the endothelial lineage. MSCs were generated out of mononuclear bone marrow cells from healthy donors separated by density gradient centrifugation. Cells were characterized by flow cytometry using a panel of monoclonal antibodies and were tested for their potential to differentiate along different mesenchymal lineages. Isolated MSCs were positive for the markers CD105, CD73, CD166, CD90, and CD44 and negative for typical hematopoietic and endothelial markers. They were able to differentiate into adipocytes and osteocytes after cultivation in respective media. Differentiation into endothelial-like cells was induced by cultivation of confluent cells in the presence of 2% fetal calf serum and 50 ng/ml vascular endothelial growth factor. Laser scanning cytometry analysis of the confluent cells in situ showed a strong increase of expression of endothelial-specific markers like KDR and FLT-1, and immunofluorescence analysis showed typical expression of the von Willebrand factor. The functional behavior of the differentiated cells was tested with an in vitro angiogenesis test kit where cells formed characteristic capillary-like structures. We could show the differentiation of expanded adult human MSCs into cells with phenotypic and functional features of endothelial cells. These predifferentiated cells provide new options for engineering of artificial tissues based on autologous MSCs and vascularized engineered tissues. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) - ReferenceVaysse L et al. (FEB 2004) The Journal of biological chemistry 279 7 5555--64
Development of a self-assembling nuclear targeting vector system based on the tetracycline repressor protein.
The ultimate destination for most gene therapy vectors is the nucleus and nuclear import of potentially therapeutic DNA is one of the major barriers for nonviral vectors. We have developed a novel approach of attaching a nuclear localization sequence (NLS) peptide to DNA in a non-essential position, by generating a fusion between the tetracycline repressor protein TetR and the SV40-derived NLS peptide. The high affinity and specificity of TetR for the short DNA sequence tetO was used in these studies to bind the NLS to DNA as demonstrated by the reduced electrophoretic mobility of the TetR.tetO-DNA complexes. The protein TetR-NLS, but not control protein TetR, specifically enhances gene expression from lipofected tetO-containing DNA between 4- and 16-fold. The specific enhancement is observed in a variety of cell types, including primary and growth-arrested cells. Intracellular trafficking studies demonstrate an increased accumulation of fluorescence labeled DNA in the nucleus after TetR-NLS binding. In comparison, binding studies using the similar fusion of peptide nucleic acid (PNA) with NLS peptide, demonstrate specific binding of PNA to plasmid DNA. However, although we observed a 2-8.5-fold increase in plasmid-mediated luciferase activity with bis-PNA-NLS, control bis-PNA without an NLS sequence gave a similar increase, suggesting that the effect may not be because of a specific bis-PNA-NLS-mediated enhancement of nuclear transfer of the plasmid. Overall, we found TetRNLS-enhanced plasmid-mediated transgene expression at a similar level to that by bis-PNA-NLS or bis-PNA alone but specific to nuclear uptake and significantly more reliable and reproducible. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human) - ReferenceTauchmanovà et al. (MAY 2003) Cancer 97 10 2453--61
Avascular necrosis in long-term survivors after allogeneic or autologous stem cell transplantation: a single center experience and a review.
BACKGROUND: The most debilitating skeletal complication of stem cell transplantation (SCT) is avascular necrosis (AVN). METHODS: Two hundred seven consecutive patients were evaluated prospectively for AVN. They survived disease free for more than 180 days after autologous or allogeneic SCT for hematologic malignancies. The diagnosis of AVN in suspicious cases was confirmed by magnetic resonance imaging. Possible correlations with treatments, bone mineral density (BMD), graft versus host disease (GVHD), and in vitro growth of fibroblast progenitors were investigated. Bone mineral density was evaluated by dual-energy X-ray absorptiometry in 100 transplanted patients, and the in vitro growth of fibroblast progenitors was monitored by a fibroblast colony-forming unit (CFU-F) assay in 30 patients after allogeneic SCT. RESULTS: Twelve patients developed AVN 3-114 months (median, 26 months) following SCT: 10 (10%) after allogeneic SCT and 2 (1.9%) after autologous SCT (P = 0.04). Twenty-five joints were affected by AVN. All patients had femoral head involvement, which was managed with hip replacement in six of them. All but one patient who developed AVN after allogeneic SCT suffered from chronic GVHD (cGVHD). Avascular necrosis occurred 1-4 months after exacerbation or progression of cGVHD. Cumulative dose of steroids was similar in both SCT groups (including steroids given pretransplant for the basic disease), whereas treatment duration was significantly longer in the allogeneic SCT group. Avascular necrosis was related to the decreased number of bone marrow CFU-F colonies in vitro, but not to BMD values. CONCLUSIONS: Avascular necrosis is a skeletal complication that occurs more often after allogeneic than after autologous SCT. Occurrence of AVN symptoms after clinical follow-up of cGVHD suggests that cGVHD requiring long-term steroid therapy is one of the main risk factors for AVN. Avascular necrosis may be facilitated by a severe deficit in the repopulating capacity of bone marrow stromal stem cells after SCT. View PublicationCatalog #: Product Name: 05401 MesenCult™ MSC Basal Medium (Human) 05402 MesenCult™ MSC Stimulatory Supplement (Human) 05411 MesenCult™ Proliferation Kit (Human) Catalog #: 05401 Product Name: MesenCult™ MSC Basal Medium (Human) Catalog #: 05402 Product Name: MesenCult™ MSC Stimulatory Supplement (Human) Catalog #: 05411 Product Name: MesenCult™ Proliferation Kit (Human)
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