Cord Blood

California institute for regenerative medicine: accelerating stem cell therapies in california and beyond.

Stem Cells. 2012 Mar;30(3):357-9

Authors: Trounson A

PMID: 22334457 [PubMed - in process]

Reduced-intensity conditioning with combined haploidentical and cord blood transplantation results in rapid engraftment, low GVHD, and durable remissions.

Blood. 2011 Dec 8;118(24):6438-45

Authors: Liu H, Rich ES, Godley L, Odenike O, Joseph L, Marino S, Kline J, Nguyen V, Cunningham J, Larson RA, del Cerro P, Schroeder L, Pape L, Stock W, Wickrema A, Artz AS, van Besien K

Abstract
We conducted a 45 patient prospective study of reduced-intensity conditioning (RIC) and transplantation of unrelated umbilical cord blood (UCB) and CD34(+) stem cells from a haploidentical family member. Median age was 50 years; weight was 80 kg. Fifty-eight percent had active disease. Neutrophil engraftment occurred at 11 days (interquartile range [IQR], 9-15) and platelet engraftment at 19 days (IQR, 15-33). In the majority of patients, early haploidentical engraftment was replaced by durable engraftment of UCB by 100 days, with regular persistence of minor host and/or haplo-hematopoiesis. Percentage of haplochimerism at day 100 correlated with the haplo-CD34 dose (P = .003). Cumulative incidence of acute GVHD (aGVHD) was 25% and chronic GVHD (cGVHD) was 5%. Actuarial survival at 1 year was 55%, progression-free survival (PFS) was 42%, nonrelapse mortality (NRM) was 28%, and relapse was 30%. RIC and haplo-cord transplantation results in fast engraftment of neutrophils and platelets, low incidences of aGVHD and cGVHD, low frequency of delayed opportunistic infections, reduced transfusion requirements, shortened length of hospital stay, and promising long-term outcomes. UCB cell dose had no impact on time to hematopoietic recovery. Therefore, UCB selection can prioritize matching, and better matched donors can be identified rapidly for most patients. This study is registered at http://clinicaltrials.gov as NCI clinical trial no. NCT00943800.

PMID: 21976674 [PubMed - indexed for MEDLINE]

Human cord blood-derived multipotent stem cells (CB-SCs) treated with all-trans-retinoic acid (ATRA) give rise to dopamine neurons.

Biochem Biophys Res Commun. 2012 Feb 4;

Authors: Li X, Li H, Bi J, Chen Y, Jain S, Zhao Y

Abstract
Parkinson's disease (PD) results from the chronic degeneration of dopaminergic neurons. A replacement for these neurons has the potential to provide a clinical cure and/or lasting treatment for symptoms of the disease. Human cord blood-derived multipotent stem cells (CB-SCs) display embryonic stem cell characteristics, including multi-potential differentiation. To explore their therapeutic potential in PD, we examined whether CB-SCs could be induced to differentiate into dopamine neurons in the presence of all-trans retinoic acid (ATRA). Prior to treatment, CB-SCs expressed mRNA and protein for the key dopaminergic transcription factors Nurr1, Wnt1, and En1. Following treatment with 10μM ATRA for 12days, CB-SCs displayed elongated neuronal-like morphologies. Immunocytochemistry revealed that 48±11% of ATRA-treated cells were positive for tyrosine hydroxylase (TH), and 36±9% of cells were positive for dopamine transporter (DAT). In contrast, control CB-SCs (culture medium only) expressed only background levels of TH and DAT. Finally, ATRA-treated CB-SCs challenged with potassium released increased levels of dopamine compared to control. These data demonstrate that ATRA induces differentiation of CB-SCs into dopaminergic neurons. This finding may lead to the development of an alternative approach to stem cell therapy for Parkinson's disease.

PMID: 22330803 [PubMed - as supplied by publisher]

Response to Comment on 'Stem-cell Therapy for Peripheral Arterial Occlusive Disease'

Eur J Vasc Endovasc Surg. 2012 Feb 9;

Authors: Kim DI

PMID: 22326694 [PubMed - as supplied by publisher]

Implications of different fluid overload definitions in pediatric stem cell transplant patients requiring continuous renal replacement therapy.

Intensive Care Med. 2012 Feb 11;

Authors: Lombel RM, Kommareddi M, Mottes T, Selewski DT, Han YY, Gipson DS, Collins KL, Heung M

Abstract
PURPOSE: In critically ill pediatric patients, fluid overload (FO) >10% has been identified as a threshold for possible interventions, including initiation of continuous renal replacement therapy (CRRT). However, multiple definitions have been reported, and there remains no consensus method for FO calculation. The goal of this study was to compare different methods of FO determination and to assess their relative value in predicting outcomes. METHODS: This is a retrospective single-center review of 21 pediatric stem cell transplant patients (PSCT) that required CRRT from 2004 to 2009. We compared eight definitions (4 weight-based and 4 fluid-balance based) that varied by baseline weights. Outcome measures were pediatric intensive care unit (PICU) mortality and pediatric logistic organ dysfunction (PELOD) scores. RESULTS: The number of patients identified as having >10% FO varied significantly according to the definition used, from 14 to 48% (p = 0.002). Significant intra-subject variability was observed; the median difference between individual minimum and maximum %FO scores was 11.4% (IQR 6.8, 17.1%). %FO was not significantly associated with PICU mortality, but five of eight FO definitions were predictive of higher subsequent PELOD scores. CONCLUSION: Our study is one of the first to compare different FO definitions and the impact on predicting outcomes. Our findings suggest that depending on the FO definition used, there is significant variability in the calculated %FO in PSCT patients, and this has important implications for clinical decision-making. Further studies are necessary to determine an optimal FO definition that is clinically relevant and predictive of important outcomes.

PMID: 22327560 [PubMed - as supplied by publisher]

Myeloablative Transplantation using either Cord Blood or Bone Marrow leads to Immune Recovery, High Long-Term Donor Chimerism and Excellent Survival in Chronic Granulomatous Disease.

Biol Blood Marrow Transplant. 2012 Feb 9;

Authors: Tewari P, Martin PL, Mendizabal A, Parikh SH, Page KM, Driscoll TA, Malech HL, Kurtzberg J, Prasad VK

Abstract
The curative potential of hematopoietic stem cell transplantation (HSCT) in patients with chronic granulomatous disease (CGD) depends upon availability of a suitable donor, successful donor engraftment and maintenance of long-term donor chimerism. Twelve consecutive children (median age 59.5 months; range, 8-140) with severe CGD (serious bacterial/fungal infections pre-transplant, median 3; range, 2-9) received myeloablative HSCT using sibling bone marrow (SibBM; n=5), unrelated cord blood (UCB; n=6), and sibling cord blood (SibCB; n=1) at our center between 1997-2010. SibBM and SibCB were HLA matched at 6/6 while UCB were 5/6 (n=5) or 6/6 (n=1). Recipients of SibBM were conditioned with Busulfan and Cyclophosphamide (Bu/Cy) ± ATG while 6 of 7 CB recipients received Fludarabine/Bu/Cy/ATG. Seven patients received G-CSF-mobilized granulocyte transfusions from directed donors. The first 2 UCB recipients had primary graft failure but were successfully re-transplanted with UCB. Highest acute GvHD was grade III (n=1). Extensive chronic GvHD developed in 3 patients. All patients are alive with median follow-up of 70.5 (range 12-167) months with high donor chimerism (>98%, n=10; 94%, n=1; and 92%, n=1). Myeloablative HSCT led to correction of neutrophil dysfunction, durable donor chimerism, excellent survival, good quality of life, and low incidence of GvHD regardless of graft source.

PMID: 22326631 [PubMed - as supplied by publisher]

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells.

Cytotechnology. 2012 Feb 11;

Authors: Bakhshandeh B, Soleimani M, Hafizi M, Ghaemi N

Abstract
The focus of both clinical and basic studies on stem cells is increasing due to their potentials in regenerative medicine and cell-based therapies. Recently stem cells have been genetically modified to enhance an existing character in or to bring a new property to them. However, accomplishment of declared goals requires detailed knowledge about their molecular characteristics which could be achieved by genetic modifications mostly through nonviral transfection strategies. Capable of differentiating into multiple cells, human unrestricted somatic stem cells (hUSSCs) and human mesenchymal stem cells (hMSCs) seem to be suitable candidates for transfection approaches. Involvement of microRNAs (miRNAs) in many biological processes makes their transfection evaluation valuable. Herein we investigated the efficacy and toxicity of four typically used transfection reagents (Arrest-In, Lipofectamine 2000, Oligofectamine and HiPerfect) systematically to deliver fluorescent labeled-miRNA and Green Fluorescent Protein (GFP) expressing plasmid into hUSSCs and hMSCs. The authenticity of stem cells was verified by differentiation experiments along with flow cytometry of surface markers. Our study revealed that stemness properties of these stem cells were not affected by transient transfection. Moreover the ratios of cell viability and transfection efficiency in both analyzed stem cells were reversed. Considering cell viability, the highest fraction of GFP-expressing cells was obtained using Oligofectamine (~50%) while the highest transfection rate of miRNA was achieved by Lipofectamine 2000 (~90%). Moreover dependency of hMSCs to size of transfected nucleic acid and time-dependency of Oligofectamine and their affection on the yield of transfection were observed. Cytotoxicity assessments also showed that hUSSCs are sensitive to HiPerFect. In addition cells treated by Lipofectamine showed morphological changes. Representing the efficient nucleic acid transfection, our research facilitates comprehensive genetic modification of stem cells and demonstrates powerful approaches to understand stem cell molecular regulation mechanisms, which eventually improves nonviral cell-mediated gene therapy.

PMID: 22328133 [PubMed - as supplied by publisher]

Mesenchymal stem cells from umbilical cord blood: parameters for isolation, characterization and adipogenic differentiation.

Cytotechnology. 2012 Feb 12;

Authors: Sibov TT, Severino P, Marti LC, Pavon LF, Oliveira DM, Tobo PR, Campos AH, Paes AT, Amaro E, F Gamarra L, Moreira-Filho CA

Abstract
Isolation of mesenchymal stem cells (MSCs) from umbilical cord blood (UCB) from full-term deliveries is a laborious, time-consuming process that results in a low yield of cells. In this study we identified parameters that can be helpful for a successful isolation of UCB-MSCs. According to our findings, chances for a well succeeded isolation of these cells are higher when MSCs were isolated from UCB collected from normal full-term pregnancies that did not last over 37 weeks. Besides the duration of pregnancy, blood volume and storage period of the UCB should also be considered for a successful isolation of these cells. Here, we found that the ideal blood volume collected should be above 80 mL and the period of storage should not exceed 6 h. We characterized UCB-MSCs by morphologic, immunophenotypic, protein/gene expression and by adipogenic differentiation potential. Isolated UCB-MSCs showed fibroblast-like morphology and the capacity of differentiating into adipocyte-like cells. Looking for markers of the undifferentiated status of UCB-MSCs, we analyzed the UCB-MSCs' protein expression profile along different time periods of the differentiation process into adipocyte-like cells. Our results showed that there is a decrease in the expression of the markers CD73, CD90, and CD105 that correlates to the degree of differentiation of UCB-MSCs We suggest that CD90 can be used as a mark to follow the differentiation commitment degree of MSCs. Microarray results showed an up-regulation of genes related to the adipogenesis process and to redox metabolism in the adipocyte-like differentiated MSCs. Our study provides information on a group of parameters that may help with successful isolation and consequently with characterization of the differentiated/undifferentiated status of UCB-MSCs, which will be useful to monitor the differentiation commitment of UCB-MSC and further facilitate the application of those cells in stem-cell therapy.

PMID: 22328147 [PubMed - as supplied by publisher]