Cord Blood

Cord Blood Stem Cell-derived DCs Generate Potent Antigen-Specific Immune Responses and Anti-tumor Effects.

Clin Sci (Lond). 2012 Jan 23;

Authors: Chang MC, Lee CN, Chen YL, Chiang YC, Sun WZ, Hu YH, Chen CA, Cheng WF

Abstract
This study aims to evaluate if cord blood stem cells (CBSCs) can be new source of dendritic cells (DCs) which can generate more potent antigen-specific immune responses and anti-tumor effects. The CBSCs and peripheral blood mononuclear cells (PBMCs) were collected, cultured and differentiated into DCs. Surface markers, secreting cytokines, antigen presentation activity, antigen-specific cell-mediated immunity and cytotoxic killing effects induced by these two origins of DCs were evaluated and compared. The CBSCs expanded for ~17-fold by ex vivo culture. The expressions of surface markers in CBSC-derived DCs were higher than those in PBMC-derived DCs treated with LPS. The CBSC-derived DCs mainly secreted IL-6, IL-10, and TNF-a, while PBMC-derived DCs mainly secreted IL-5 and IFN-γ. The CBSC-derived DCs had better antigen presentation abilities when stimulated with LPS or TNF-a, induced higher numbers of IFN-g-secreting, antigen-specific CD8+ T lymphocytes by ELLIspot assay, and stimulated stronger antigen-specific CTL activities (p<0.01, one-way ANOVA). The CBSC-derived DCs showed quicker and stronger ERK and Akt phosphorylation, and weaker p38 phosphorylation than PBMC-derived DCs when stimulated with LPS. The CBSC-derived DCs have abilities of inducing stronger antigen-specific immunity and more potent anti-tumor effects. The CBSCs can be a good source of DCs in the strategy of DC-based cancer vaccine and immunotherapy.

PMID: 22264240 [PubMed - as supplied by publisher]

Impact of DAG stimulation on mineral synthesis, mineral structure and osteogenic differentiation of human cord blood stem cells.

Stem Cell Res. 2012 Mar;8(2):193-205

Authors: Lammers L, Naujoks C, Berr K, Depprich R, Kübler N, Meyer U, Langenbach F, Lüttenberg B, Kögler G, Wiesmann HP, Handschel J

Abstract
It remains unexplored in what way osteogenic stimulation with dexamethasone, ascorbic acid and β-glycerol phosphate (DAG) influences the process of mineralization, the composition and structure of the assembled mineral. Therefore, we analyzed and characterized biomineralization in DAG-stimulated and unstimulated 3D human unrestricted somatic stem cell (USSC) cultures. Microspheres were analyzed by histological staining, scanning electron microscopy (SEM), semi-quantitative energy-dispersive X-ray spectroscopy (EDX), quantitative wavelength-dispersive X-ray spectroscopy (WDX), transmission electron microscopy (TEM), selected area electron diffraction (SAED) and Raman spectroscopy. Mineral material was detected by SEM and histological staining in both groups, and showed structural differences. DAG influenced the differentiation of USSCs and the formation, structure and composition of the assembled mineral. SEM showed that cells of the +DAG spheres exhibited morphological signs of osteoblast-like cells. EDX and WDX confirmed a Ca-P mineral in both groups. Overall, the mineral material found showed structural similarities to the mineral substance of bony material.

PMID: 22265739 [PubMed - in process]

Hypoxia improves expansion potential of human cord blood-derived hematopoietic stem cells and marrow repopulation efficiency.

Eur J Haematol. 2012 Jan 23;

Authors: Roy S, Tripathy M, Mathur N, Jain A, Mukhopadhyay A

Abstract
Objectives:  In bone marrow hematopoietic stem cells (HSCs) reside in the most hypoxic endosteum niche; whereas, the proliferating progenitors are located near the relatively oxygen rich vascular region. High oxygen tension is potentially detrimental to HSCs. The objective of this investigation was to compare cellular, functional and molecular responses of human umbilical cord blood (UCB)-derived hematopoietic stem and progenitor cells in culture under hypoxic and normoxic conditions. Methods:  CD133-enriched UCB cells were cultured in growth factor containing serum-free and serum-supplemented medium under 5% O(2) (hypoxia) or 21% O(2) (normoxia) for 10 days. The phenotypes of expanded cells were analyzed by flowcytometry and the engraftability by SCID-repopulation assay. The expression of hypoxia-inducible factor (HIF)-1α and some of its target genes were analyzed by real-time RT-PCR. Results:  In hypoxic culture, CD34(+) CD38(-) cells were expanded about 27-fold, which was significantly (p < 0.01) higher than that obtained in normoxic culture. Serum free culture did not support the growth of cells in the presence of 21% O(2) . Myeloid colony forming potential of cells was significantly (p < 0.05) increased in 5% O(2) that to 21% O(2) culture. SCID-repopulation efficiency seems to be better preserved in the cells cultured under hypoxic conditions. Hypoxia significantly (p < 0.05) induced the expression of HIF-1α, vascular endothelial growth factor (VEGF) and ABCG2 genes, and also upregulated CXCR4 receptor expression. Conclusions:  Low oxygen tension enhanced the proliferation of UCB-derived HSC/progenitor cells and maintenance of SRCs than normoxia. These expanded cells are expected to be beneficial in the patients who lack human leukocyte antigen (HLA)-matched donors. © 2012 John Wiley & Sons A/S.

PMID: 22268587 [PubMed - as supplied by publisher]

Isolation, characterization and differentiation of mesenchymal stem cells (MSCs) from amniotic fluid, cord blood and Wharton's jelly in the horse.

Reproduction. 2012 Jan 24;

Authors: Iacono E, Brunori L, Pirrone A, Ricci F, Pagliaro PP, Tazzari PL, Merlo B

Abstract
Mesenchymal stem cells have been derived from multiple sources in the horse including umbilical cord blood and amnion. This work aimed to identify and characterize stem cells from equine amniotic fluid (AF), cord blood (CB) and Wharton's jelly (WJ). Samples were recovered from 13 mares at labour. AF and CB cells were isolated by centrifugation, while WJ was incubated with an enzimatic solution for 2 h. All cell lines were cultured in DMEM/TCM199 plus FBS. Fibroblast-like cells were observed in 7/10 (70%) AF, 6/8 (75%) CB, and 8/12 (66.7%) WJ samples. Statistically significant differences were found between doubling times: cells isolated from WJ expanded more rapidly (2.0±0.6 days) than those isolated from CB (2.6±1.3 days) and AF (2.3±1.0 days) (P<0.05). Positive von Kossa and Alizarin Red staining confirmed osteogenesis. Alcian blue staining of matrix glycosaminoglycans illustrated chondrogenesis, and positive Oil Red O lipid droplets staining suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105 and negative for CD34, CD14 and CD45. These findings suggest that equine MSCs from AF, UCB and WJ appeared to be a readily obtainable and highly proliferative cell lines from an uninvasive source that may represent a good model system for stem cell biology and cellular therapy applications in horses. However, to assess their usefulness as an allogenic cell source, further studies are needed for evaluating expression of markers related to cell immunogenicity.

PMID: 22274885 [PubMed - as supplied by publisher]

High-dose therapy with autologous stem cell transplantation versus chemotherapy or immuno-chemotherapy for follicular lymphoma in adults.

Cochrane Database Syst Rev. 2012;1:CD007678

Authors: Schaaf M, Reiser M, Borchmann P, Engert A, Skoetz N

Abstract
BACKGROUND: Follicular lymphoma (FL) is the most common indolent and second most common Non-Hodgkin`s lymphoma (NHL) in the Western world. Standard treatment usually includes rituximab and chemotherapy. High-dose therapy (HDT) followed by autologous stem cell transplantation (ASCT) is an option for patients in advanced stages or for second-line therapy, leading to improved progression-free survival (PFS) rates. However, the impact of HDT and ASCT remains unclear, as there are hints of an increased risk of second cancers.
OBJECTIVES: We performed a systematic review with meta-analysis of randomised controlled trials (RCTs) comparing HDT plus ASCT with chemotherapy or immuno-chemotherapy in patients with FL with respect to overall survival (OS), PFS, treatment-related mortality (TRM), adverse events and secondary malignancies.
SEARCH METHODS: We searched CENTRAL, MEDLINE, and EMBASE as well as conference proceedings from January 1985 to September 2011 for RCTs. Two review authors independently screened search results.
SELECTION CRITERIA: Randomised controlled trials comparing chemotherapy or immuno-chemotherapy with HDT followed by ASCT in adults with previously untreated or relapsed FL.
DATA COLLECTION AND ANALYSIS: We used hazard ratios (HR) as effect measures used for OS and PFS as well as relative risks for response rates. Two review authors independently extracted data and assessed the quality of trials.
MAIN RESULTS: Our search strategies led to 3046 potentially relevant references. Of these, five RCTs involving 1093 patients were included; four trials in previously untreated patients and one trial in relapsed patients. Overall, the quality of the five trials is judged to be moderate. All trials were reported as randomised and judged to be open-label studies, because usually trials evaluating stem cell transplantation are not blinded. Due to the small number of studies in each analysis (four or less), the quantification of heterogeneity was not reliable and not evaluated in further detail. A potential source of bias are uncertainties in the HR calculation. For OS, the HR had to be calculated for three trials from survival curves, for PFS for two trials.We found a statistically significant increased PFS in previously untreated FL patients in the HDT + ASCT arm (HR = 0.42 (95% confidence interval (CI) 0.33 to 0.54; P < 0.00001). However, this effect is not transferred into a statistically significant OS advantage (HR = 0.97; 95% 0.76 to 1.24; P = 0.81). The subgroup of trials adding rituximab to both intervention arms (one trial) confirms these results and the trial had to be stopped early after an interim analysis due to a statistically significant PFS advantage in the HDT + ASCT arm (PFS: HR = 0.36; 95% CI 0.23 to 0.55; OS: HR = 0.88; 95% CI 0.40 to 1.92). In the four trials in previously untreated patients there are no statistically significant differences between HDT + ASCT and the control-arm in terms of TRM (RR = 1.28; 95% CI 0.25 to 6.61; P = 0.77), secondary acute myeloid leukaemia/myelodysplastic syndromes (RR = 2.87; 95% CI 0.7 to 11.75; P = 0.14) or solid cancers (RR = 1.20; 95% CI 0.25 to 5.77; P = 0.82). Adverse events were rarely reported and were observed more frequently in patients undergoing HDT + ASCT (mostly infections and haematological toxicity).For patients with relapsed FL, there is some evidence (one trial, N = 70) that HDT + ASCT is advantageous in terms of PFS and OS (PFS: HR = 0.30; 95% CI 0.15 to 0.61; OS: HR = 0.40; 95% CI 0.18 to 0.89). For this trial, no results were reported for TRM, adverse events or secondary cancers.
AUTHORS' CONCLUSIONS: In summary, the currently available evidence suggests a strong PFS benefit for HDT + ASCT compared with chemotherapy or immuno-chemotherapy in previously untreated patients with FL. No statistically significant differences in terms of OS, TRM and secondary cancers were detected. These effects are confirmed in a subgroup analysis (one trial) adding rituximab to both treatment arms. Further trials evaluating this approach are needed to determine this effect more precisely in the era of rituximab. Moreover, longer follow-up data are necessary to find out whether the PFS advantage will translate into an OS advantage in previously untreated patients with FL.There is evidence that HDT + ASCT is advantageous in patients with relapsed FL.

PMID: 22258971 [PubMed - in process]

Allogeneic stem cell transplantation versus conventional therapy for advanced primary cutaneous T-cell lymphoma.

Cochrane Database Syst Rev. 2012;1:CD008908

Authors: Schlaak M, Pickenhain J, Theurich S, Skoetz N, von Bergwelt-Baildon M, Kurschat P

Abstract
BACKGROUND: Primary cutaneous T-cell lymphomas (CTCL) belong to the group of non-Hodgkin lymphomas and usually run an indolent course. However, some patients progress to advanced tumour or leukaemic stages. Up to now, no curative treatment has been established for those cases. In the last few years, several publications have reported durable responses in some patients following allogeneic stem cell transplantation (alloSCT).
OBJECTIVES: To compare the efficacy and safety of conventional therapies with allogeneic stem cell transplantation in patients with advanced primary cutaneous T-cell lymphomas.
SEARCH METHODS: The search strategy included the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE (1950 to May 2011), Internet-databases of ongoing trials (www.controlled-trials.com; www.clinicaltrials.gov), conference proceedings of the American Society of Clinical Oncology (ASCO, 2009 to present) and the American Society of Hematology (ASH, 2009 to present). We also contacted members of the European Organisation for Research and Treatment of Cancer (EORTC) Cutaneous Lymphoma Task Force to check for ongoing study activities. We handsearched citations from identified trials and relevant review articles. In addition, randomised controlled trials from the European Group for Blood and Marrow Transplantation (EBMT) and International Conference on Cutaneous T-cell Lymphoma, ASCO and ASH up to 2010 were handsearched.
SELECTION CRITERIA: Genetically randomised controlled trials (RCT) comparing alloSCT plus conditioning therapy regardless of agents with conventional therapy as treatment for advanced CTCL were eligible to be included.
DATA COLLECTION AND ANALYSIS: From eligible studies data would have been extracted by two review authors and assessed for quality. Primary outcome measures were overall survival, secondary criteria were time to progression, response rate, treatment-related mortality, adverse events and quality of life.
MAIN RESULTS: We found 2077 citations but none were relevant genetically or non-genetically randomised controlled trials. All 41 studies that were thought to be potentially suitable were excluded after full text screening for being non-randomised, not including CTCL or being review articles.
AUTHORS' CONCLUSIONS: We planned to report evidence from genetically or non-genetically randomised controlled trials comparing conventional therapy and allogeneic stem cell transplantation. However, no randomised trials addressing this question were identified. Nevertheless, prospective genetically randomised controlled trials need to be initiated to evaluate the precise role of alloSCT in advanced CTCL.

PMID: 22258991 [PubMed - in process]

Cord blood meets its match.

Lancet Oncol. 2011 Dec;12(13):1177-8

Authors: Gratwohl A

PMID: 21982421 [PubMed - indexed for MEDLINE]

Polyunsaturated fatty acids confer cryoresistance on megakaryocytes generated from cord blood and also enhance megakaryocyte production from cryopreserved cord blood cells.

Cytotherapy. 2012 Jan 17;

Authors: Shabrani NC, Khan NF, Kale VP, Limaye LS

Abstract
Background aims. Previous data have shown that the addition of docosahexanoic acid (DHA)/arachidonic acid (AA) has a beneficial effect on cytokine-mediated in vitro generation of megakaryocytes (MK) from umbilical cord blood (UCB).Cryopreservation forms an inherent part of UCB banking and MK progenitors are known to be very sensitive to the stresses of freezing. It is therefore imperative to generate functional cells from cryopreserved cells, and the generated cells need to be cryopreserved until used. In the present study, cryopreservation of ex vivo-expanded MK as well as MK generation from cryopreserved UCB samples was investigated. Methods. MK generated with or without DHA/AA were cryopreserved in freezing medium containing 10% dimethyl sulfoxide (DMSO). Freezing efficacy was tested by quantitating MK after revival. Cryopreserved CD34(+) cells were cultured with stem cell factor (SCF) and thrombopoietin (TPO), in the presence and absence of DHA/AA for 10 days, and then quantitated for MK. Results. We observed a 1.5-3-fold increase in MK numbers, their progenitor content and their expression of phenotypic markers and MK-related transcription factors. DHA/AA sets showed a 2-5-fold improved engraftment in NOD/SCID mice. These data showed that the beneficial effect of DHA/AA obtained during MK expansion was not altered after freezing stress. The enhancement in MK generation obtained from fresh cord blood (CB) cells was reproduced with comparable efficiency when we used cryopreserved CB samples. Conclusions. Taken together, our data suggest that in vitro-generated DHA/AA MK survive cryoinjuries in a functionally better state. DHA/AA support a more efficient generation of MK from cryopreserved UCB.

PMID: 22250991 [PubMed - as supplied by publisher]