A higher CD34 + cell dose correlates with better event-free survival after KIR-ligand mismatched cord blood transplantation for childhood acute myeloid leukemia.

Although killer Ig-like receptor ligands (KIR-L) mismatch has been associated with alloreactive natural killer cell activity and potent graft-versus-leukemia (GVL) effect among adults with acute myeloid leukemia (AML), its role among children with AML receiving cord blood transplantation (CBT) has not been determined. We conducted a retrospective study using a nationwide registry of the Japanese Society for Transplantation and Cellular Therapy. Patients who were diagnosed with de novo non-M3 AML and who underwent their first CBT in remission between 2000 and 2021 at under 16 years old were included. A total of 299 patients were included; 238 patients were in the KIR-L match group, and 61 patients were in the KIR-L mismatch group. The cumulative incidence rates of neutrophil recovery, platelet engraftment, and acute/chronic graft-versus-host disease did not differ significantly between the groups. The 5-year event-free survival (EFS) rate was 69.8% in the KIR-L match group and 74.0% in the KIR-L mismatch group (p = 0.490). Stratification by CD34 + cell dose into four groups revealed a significant correlation between CD34 + cell dose and EFS in the KIR-L mismatch group (p = 0.006) but not in the KIR-L match group (p = 0.325). According to our multivariate analysis, KIR-L mismatch with a high CD34 + cell dose (≥ median dose) was identified as an independent favorable prognostic factor for EFS (hazard ratio = 0.19, p = 0.029) and for the cumulative incidence of relapse (hazard ratio = 0.09, p = 0.021). Our results suggested that higher CD34 + cell doses are crucial for achieving a potent GVL effect in the context of KIR-L-mismatched CBT.

Role of Nuclear-Receptor-Related 1 in the Synergistic Neuroprotective Effect of Umbilical Cord Blood and Erythropoietin Combination Therapy in Hypoxic Ischemic Encephalopathy.

Neonatal hypoxic-ischemic encephalopathy (HIE) results in neurological impairments; cell-based therapy has been suggested as a therapeutic avenue. Previous research has demonstrated the synergistically potentiated therapeutic efficacy of human umbilical cord blood (UCB) by combining recombinant human erythropoietin (EPO) treatment for recovery from HIE. However, its molecular mechanism is not entirely understood. In the present study, we analyzed the mechanisms underlying the effect of combination treatment with EPO and UCB by transcriptomic analysis, followed by gene enrichment analysis. Mouse HIE model of the neonate was prepared and randomly divided into five groups: sham, HIE, and UCB, EPO, and UCB+EPO treatments after HIE. A total of 376 genes were differentially expressed when |log2FC| ≥ 1-fold change expression values were considered to be differentially expressed between UCB+EPO and HIE. Further assessment through qRT-PCR and gene enrichment analysis confirmed the expression and correlation of its potential target, Nurr1, as an essential gene involved in the synergistic effect of the UCB+EPO combination. The results indicated the remarkable activation of Wnt/ß-catenin signaling by reducing the infarct size by UCB+EPO treatment, accompanied by Nurr1 activity. In conclusion, these findings suggest that the regulation of Nurr1 through the Wnt/ß-catenin pathway exerts a synergistic neuroprotective effect in UCB and EPO combination treatment.

Synergistic Effect in Neurological Recovery via Anti-Apoptotic Akt Signaling in Umbilical Cord Blood and Erythropoietin Combination Therapy for Neonatal Hypoxic-Ischemic Brain Injury.

Our previous clinical studies demonstrated the synergistic therapeutic effect induced by co-administering recombinant human erythropoietin (rhEPO) in human umbilical cord blood (hUCB) therapy for children with cerebral palsy. However, the cellular mechanism beyond the beneficial effects in this combination therapy still needs to be elucidated. A hypoxic-ischemic encephalopathy (HIE) model of neonates, representing cerebral palsy, was prepared and randomly divided into five groups (hUCB+rhEPO combination, hUCB, and rhEPO treatments over HIE, HIE control, and sham). Seven days after, hUCB was administered intraperitoneally and the rhEPO injections were started. Neurobehavioral tests showed the best outcome in the combination therapy group, while the hUCB and rhEPO alone treatments also showed better outcomes compared with the control (p < 0.05). Inflammatory cytokines were downregulated by the treatments and attenuated most by the combination therapy (p < 0.05). The hUCB+rhEPO treatment also showed remarkable increase in phosphorylation of Akt and potentiation of anti-apoptotic responses with decreased Bax and increased Bcl-2 (p < 0.05). Pre-treatment of MK-2206, an Akt inhibitor, for the combination therapy depressed the anti-apoptotic effects. In conclusion, these findings suggest that the therapeutic effect of hUCB therapy might be potentiated by co-administration of rhEPO via augmentation of anti-inflammatory and anti-apoptotic responses related to the phosphorylation of Akt.

Efficacy of Cord Blood Cell Therapy for Hutchinson-Gilford Progeria Syndrome-A Case Report.

Hutchinson-Gilford progeria syndrome (HGPS) is an extremely rare premature aging disorder characterized by short stature and atherosclerosis-induced death within teenage years. A 13-year-old male diagnosed with HGPS was administered three intravenous infusions of allogeneic cord blood (CB) cells from unrelated donors at four-month intervals to evaluate the safety and its therapeutic efficacy. Adverse events were monitored in addition to height, weight, laboratory blood tests, joint range of motion (ROM), and carotid Doppler. Cytokine and receptor assays were also performed. The patient exhibited an increase in growth rate for both height and weight. One year after therapy initiation, evident amelioration in pulse wave velocity, bilateral maximal intima-media thickness, and dyslipidemic status were observed, which were in abrupt aggravation prior to treatment. Further, an increase in flexibility occurred in some joints of the upper extremities. No serious adverse events were observed throughout the study period and one year beyond. A molecular assay revealed downregulation of proinflammatory and atherosclerosis, representing cytokine expressions following the administration of CB cells. This is the first reported case of an allogeneic CB trial in a patient with HGPS showing therapeutic effects of CB with improvements in anthropometric measures, joint ROM with amelioration of atherosclerosis, and dyslipidemia induced by anti-inflammatory and anti-atherosclerotic responses.

Serum Cytokine Profile, Beta-Hexosaminidase A Enzymatic Activity and GM2 Ganglioside Levels in the Plasma of a Tay-Sachs Disease Patient after Cord Blood Cell Transplantation and Curcumin Administration: A Case Report.

Tay-Sachs disease (TSD) is a progressive neurodegenerative disorder that occurs due to a deficiency of a ß hexosaminidase A (HexA) enzyme, resulting in the accumulation of GM2 gangliosides. In this work, we analyzed the effect of umbilical cord blood cell transplantation (UCBCT) and curcumin administration on the course of the disease in a patient with adult TSD. The patient's serum cytokine profile was determined using multiplex analysis. The level of GM2 gangliosides in plasma was determined using mass spectrometry. The enzymatic activity of HexA in the plasma of the patient was assessed using a fluorescent substrate assay. The HexA α-subunit (HexA) concentration was determined using ELISA. It was shown that both UCBCT and curcumin administration led to a change in the patient's cytokine profile. The UCBCT resulted in an increase in the concentration of HexA in the patient's serum and in an improvement in the patient's neurological status. However, neither UCBCT nor curcumin were able to alter HexA activity and the level of GM2 in patient's plasma. The data obtained indicate that UCBCT and curcumin administration can alter the immunity of a patient with TSD, reduce the level of inflammatory cytokines and thereby improve the patient's condition.

The effect of human umbilical cord blood-derived mesenchymal stem cell media containing serum on recovery after laser treatment: A double-blinded, randomized, split-face controlled study.

BACKGROUND: Ablative CO2 fractional laser (AFL) is a common cosmetic procedure to improve skin laxity. However, due to prolonged downtime and the risk of postinflammatory hyperpigmentation, laser-assisted delivery of active ingredients as post-laser treatment has gained interest in past years. Among various active ingredients, human umbilical cord blood-derived mesenchymal stem cells (hUCBMSCs) can be a promising agent promoting skin regeneration. AIMS: We evaluated the efficacy and safety of a human cord blood cell-conditioned media containing serum and cream on patients who underwent AFL treatment. A randomized, investigator-blinded, prospective, split-face comparison study was conducted. MATERIALS AND METHODS: Twenty-three patients who underwent AFL on both cheeks applied a human umbilical cord blood-derived mesenchymal stem cell (hUCBMSC)-conditioned media containing cream with or without stem cell containing serum application. As a primary outcome measure, we evaluated the total area of microcrusts and post-treatment erythema using digital photographs. Additionally, skin biophysical parameters (corneometer, TEWL) and global improvement scores for skin texture were assessed. RESULTS: The area of total microcrusts was reduced in the study group which applied both serum and cream. The global improvement score of the post-treatment erythema was significantly reduced. Investigator-assessed global improvement scores were higher in the combination treatment group. Additionally, there was no adverse event, which was associated with the use of either hUCBMSCs containing serum or cream. CONCLUSION: The application of human cord blood cell containing serum and cream resulted in accelerated wound healing and reduced post-treatment erythema, which effectively reduced recovery time after ablative laser treatment.

Combining Human Umbilical Cord Blood Cells With Erythropoietin Enhances Angiogenesis/Neurogenesis and Behavioral Recovery After Stroke.

Disruption of blood flow in the brain induces stroke, the leading cause of death and disability worldwide. However, so far the therapeutic options are limited. Thus, the therapeutic efficacy of cell-based approaches has been investigated to develop a potential strategy to overcome stroke-induced disability. Human umbilical cord blood cells (hUCBCs) and erythropoietin (EPO) both have angiogenic and neurogenic properties in the injured brain, and their combined administration may exert synergistic effects during neurological recovery following stroke. We investigated the therapeutic potential of hUCBC and EPO combination treatment by comparing its efficacy to those of hUCBC and EPO alone. Adult male Sprague-Dawley rats underwent transient middle cerebral artery occlusion (MCAO). Experimental groups were as follows: saline (injected once with saline 7 d after MCAO); hUCBC (1.2 × 107 total nucleated cells, injected once via the tail vein 7 d after MCAO); EPO (500 IU/kg, injected intraperitoneally for five consecutive days from 7 d after MCAO); and combination of hUCBC and EPO (hUCBC+EPO). Behavioral measures (Modified Neurological Severity Score [mNSS] and cylinder test) were recorded to assess neurological outcomes. Four weeks after MCAO, brains were harvested to analyze the status of neurogenesis and angiogenesis. In vitro assays were also conducted using neural stem and endothelial cells in the oxygen-glucose deprivation condition. Performance on the mNSS and cylinder test showed the most improvement in the hUCBC+EPO group, while hUCBC- and EPO-alone treatments showed superior outcomes relative to the saline group. Neurogenesis and angiogenesis in the cortical region was the most enhanced in the hUCBC+EPO group, while the findings in the hUCBC and EPO treatment alone groups were better than those in the saline group. Astrogliosis in the brain tissue was reduced by hUCBC and EPO treatment. The reduction was largest in the hUCBC+EPO group. These results were consistent with in vitro assessments that showed the strongest neurogenic and angiogenic effect with hUCBC+EPO treatment. This study demonstrates that combination therapy is more effective than single therapy with either hUCBC or EPO for neurological recovery from subacute stroke. The common pathway underlying hUCBC and EPO treatment requires further study.

Human Cord Blood Serum-Derived APP α-Secretase Cleavage Activity is Mediated by C1 Complement.

Alzheimer's Disease (AD) is the leading cause of dementia in the elderly. In healthy individuals, amyloid precursor protein (APP) is cleaved by α-secretase, generating soluble α-amyloid precursor protein (sAPPα), which contributes neuroprotective functions in the neuronal environment. In contrast, in the neurodegenerative environment of AD patients, amyloid-ß-peptide (Aß) of either 40 or 42 residues are generated by increased activity of ß- and γ-secretase. These proteins amalgamate in specific regions of the brain, which disrupts neuronal functions and leads to cognitive impairment. Human umbilical cord blood cells (HUCBC) have proven useful as potential immunomodulatory therapies in various models of neurodegenerative diseases, including AD. Our most recent work studied the impact of umbilical cord blood serum (CBS) on modulation of sAPPα production. Heat-sensitive CBS significantly promoted sAPPα production, indicating that heat-sensitive factor(s) play(s) a role in this process. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis was used to determine the molecular source of α-secretase in purified CBS and aged blood serum (AgBS) fraction. Of the proteins identified, the subunits of C1 complex (C1q, C1r, and C1s) and alpha-2-macroglobulin showed significantly greater levels in purified α-CBS fraction (α-CBSF) compared with the AgBS fraction (AgBSF). Specifically, C1 markedly increased sAPPα and alpha-carboxyl-terminal fragment (α-CTF) production in a dose-dependent fashion, whereas C1q alone only minimally increased and C3 did not increase sAPPα production in the absence of sera. Furthermore, C1q markedly increased sAPPα and α-CTF, while decreasing Aß, in CHO/APPwt cells cultured in the presence of whole sera. These results confirm our initial assumption that APP α-secretase activity in human blood serum is mediated by complement C1, opening a potential therapeutic modality for the future of AD.

Human Umbilical Cord Blood Serum-derived α-Secretase: Functional Testing in Alzheimer's Disease Mouse Models.

Alzheimer's disease (AD) is an age-related disorder that affects cognition. Our previous studies showed that the neuroprotective fragment of amyloid procurer protein (APP) metabolite, soluble APPα (sAPPα), interferes with ß-site APP-cleaving enzyme 1 (BACE1, ß-secretase) cleavage and reduces amyloid-ß (Aß) generation. In an attempt to identify approaches to restore sAPPα levels, we found that human cord blood serum (CBS) significantly promotes sAPPα production compared with adult blood serum (ABS) and aged blood serum (AgBS) in Chinese hamster ovary cells stably expressing wild-type human APP. Interestingly, CBS selectively mediated the α-secretase cleavage of human neuron-specific recombinant APP695 in a cell-free system independent of tumor necrosis factor-α converting enzyme (TACE; a disintegrin and metalloproteinase domain-containing protein 17 [ADAM17]) and ADAM. Subsequently, using 3-step chromatographic separation techniques (i.e., diethylaminoethanol, size-exclusion, and ion-exchange chromatography), we purified and ultimately identified a CBS-specific fraction with enhanced α-secretase catalytic activity (termed αCBSF) and found that αCBSF has more than 3,000-fold increased α-secretase catalytic activity compared with the original pooled CBS. Furthermore, intracerebroventricular injection of αCBSF markedly increased cerebral sAPPα levels together with significant decreases in cerebral Aß production and abnormal tau (Thr231) phosphorylation compared with the AgBS fraction with enhanced α-secretase activity (AgBSF) treatment in triple transgenic Alzheimer's disease (3xTg-AD) mice. Moreover, AgBSF administered intraperitoneally to transgenic mice with five familial Alzheimer's disease mutations (5XFAD) via an osmotic mini pump for 6 weeks (wk) ameliorated ß-amyloid plaques and reversed cognitive impairment measures. Together, our results propose the necessity for further study aimed at identification and characterization of α-secretase in CBS for novel and effective AD therapy.

Myelodysplastic syndrome with myelofibrosis in which azacitidine therapy was effective and cord blood transplantation was carried out.

Myelodysplastic syndrome with myelofibrosis (MDS-F) is a disease with a poor prognosis, and patients with this condition are at an increased risk of engraftment failures after allogeneic hematopoietic stem cell transplantation (SCT). Azacitidine (AZA) is effective in high-risk MDS patients. However, the effects of AZA on MDS-F have not been elucidated. AZA was administered to a 62-year-old male with MDS-F for 7 days at a dose of 75 mg/m2. Hematological improvements were observed after only 1 course of treatment. No suitable donor was found through the Japan Marrow Donor Program; therefore, the patient underwent umbilical cord blood transplant (UCBT). Neutrophil engraftment was observed on day 21 after the transplant procedure. He developed acute graft versus host disease (GVHD) of the skin (stage 3/grade II), but it could be controlled using prednisolone. Chronic GVHD was not observed and he was discharged in good general condition on day 68. While treatment prior to allogeneic SCT of MDS-F has not been established, in the present case, the hematological improvement brought about by AZA likely contributed to the patient's positive response to UCBT.

Intraarterial transplantation of human umbilical cord blood mononuclear cells in hyperacute stroke improves vascular function.

BACKGROUND: Human umbilical cord blood (hUCB) cell therapy is a promising treatment for ischemic stroke. The effects of hyperacute stem cell transplantation on cerebrovascular function in ischemic stroke are, however, not well understood. This study evaluated the effects of hyperacute intraarterial transplantation of hUCB mononuclear cells (MNCs) on cerebrovascular function in stroke rats using serial magnetic resonance imaging (MRI). METHODS: HUCB MNCs or vehicle were administered to stroke rats via the internal carotid artery immediately after reperfusion at 60 min following ischemia onset. Lesion volumes were longitudinally evaluated by MRI on days 0, 2, 14, and 28 after stroke, accompanied by behavioral tests. Cerebral blood flow (CBF) and cerebrovascular reactivity were measured by perfusion MRI and CO2 functional MRI (fMRI) at 28 days post-stroke; corresponding vascular morphological changes were also detected by immunohistology in the same animals. RESULTS: We found that CBF to the stroke-affected region at 28 days was improved (normalized CBF value: 1.41 ± 0.30 versus 0.49 ± 0.07) by intraarterial transplantation of hUCB MNCs in the hyperacute stroke phase, compared to vehicle control. Cerebrovascular reactivity within the stroke-affected area, measured by CBF fMRI, was also increased (35.2 ± 3.5% versus 12.8 ± 4.3%), as well as the corresponding cerebrovascular density. Some engrafted cells appeared with microvascular-like morphology and stained positive for von Willebrand Factor (an endothelial cell marker), suggesting they differentiated into endothelial cells. Some engrafted cells also connected to host endothelial cells, suggesting they interacted with the host vasculature. Compared to the vehicle group, infarct volume at 28 days in the stem cell treated group was significantly smaller (160.9 ± 15.7 versus 231.2 ± 16.0 mm3); behavioral deficits were also markedly reduced by stem cell treatment at day 28 (19.5 ± 1.0% versus 30.7 ± 4.7% on the foot fault test; 68.2 ± 4.6% versus 86.6 ± 5.8% on the cylinder test). More tissue within initial perfusion-diffusion mismatch was rescued in the treatment group. CONCLUSIONS: Intraarterial hUCB MNC transplantation during the hyperacute phase of ischemic stroke improved cerebrovascular function and reduced behavioral deficits and infarct volume.

The effect of the colostral cells on gene expression of cytokines in cord blood cells.

Beneficial effect of maternal milk is acknowledged, but there is still question whether maternal milk from allergic mother is as good as from healthy one. In our study, we have assayed the effect of cells from colostrum of healthy and allergic mothers on gene expression of cytokines in cord blood cells of newborns of healthy and allergic mothers. Cytokines typical for Th1 (IL-2, IFN-gamma), Th2 (IL-4, IL-13), Tregs (IL-10, TGF-beta), and IL-8 were followed. We were not able to detect significant influence of colostral cells on gene expression of cytokines in cord blood after 2-day coculture using Transwell system. There was no difference in gene expression of cytokines in nonstimulated cord blood cells of newborns of healthy and allergic mothers, but generally increased gene expression of cytokines except IL-10 and TGF-beta after polyclonal stimulation was detected in cord blood cells of children of allergic mothers. There was no difference in IL-10 expression in stimulated cord blood cells of children of healthy and allergic mothers. Gene expression of TGF-beta was even decreased in stimulated cord blood cells of children of allergic mothers in comparison to healthy ones. We have not observed difference in the capacity of colostral cells of healthy and allergic mothers to influence gene expression of cytokines in cord blood cells, but we have described difference in the reactivity of cord blood cells between children of allergic and healthy mothers.

MiR-126 Contributes to Human Umbilical Cord Blood Cell-Induced Neurorestorative Effects After Stroke in Type-2 Diabetic Mice.

Diabetes mellitus (DM) is a high risk factor for stroke and leads to more severe vascular and white-matter injury than stroke in non-DM. We tested the neurorestorative effects of delayed human umbilical cord blood cell (HUCBC) treatment of stroke in type-2 diabetes (T2DM). db/db-T2DM and db/+-non-DM mice were subjected to distal middle cerebral artery occlusion (dMCAo) and were treated 3 days after dMCAo with: (a) non-DM + Phosphate buffered saline (PBS); (b) T2DM + PBS; (c) T2DM + naïve-HUCBC; (d) T2DM + miR-126(-/-) HUCBC. Functional evaluation, vascular and white-matter changes, neuroinflammation, and miR-126 effects were measured in vivo and in vitro. T2DM mice exhibited significantly decreased serum and brain tissue miR-126 expression compared with non-DM mice. T2DM + HUCBC mice exhibited increased miR-126 expression, increased tight junction protein expression, axon/myelin, vascular density, and M2-macrophage polarization. However, decreased blood-brain barrier leakage, brain hemorrhage, and miR-126 targeted gene vascular cell adhesion molecule-1 and monocyte chemotactic protein 1 expression in the ischemic brain as well as improved functional outcome were present in HUCBC-treated T2DM mice compared with control T2DM mice. MiR-126(-/-) HUCBC-treatment abolished the benefits of naïve-HUCBC-treatment in T2DM stroke mice. In vitro, knock-in of miR-126 in primary cultured brain endothelial cells (BECs) or treatment of BECs with naïve-HUCBCs significantly increased capillary-like tube formation, and increased axonal outgrowth in primary cultured cortical neurons; whereas treatment of BECs or cortical neurons with miR-126(-/-) HUCBC attenuated HUCBC-treatment-induced capillary tube formation and axonal outgrowth. Our data suggest delayed HUCBC-treatment of stroke increases vascular/white-matter remodeling and anti-inflammatory effects; MiR-126 may contribute to HUCBC-induced neurorestorative effects in T2DM mice.

Correlación entre características físicas neonatales y maternas con el recuento total de células nucleadas y de células cd34+ por microlitro en sangre de cordón umbilical / Correlation between neonatal and maternal physical characteristics with total nucleated cell count and cell count per microliter cd34+ in cord blood / Correlação entre características físicas neonatais e maternas com a contagem total de células nucleadas e células cd34 + por microlitro no sangue do cordão umbilical

Introducción: La sangre de cordón umbilical (SCU) ha sido reconocida como una fuente de células madre hematopoyéticas. Múltiples estudios han sido realizados con el propósito de determinar variables maternas y neonatales que afecten el volumen, el recuento total de células nucleadas y de células CD34+. El presente estudio correlaciona variables maternas y neonatales con el recuento total de células nucleadas y de CD34+ medidas en μL (microlitro). Materiales y Métodos: Estudio correlacional en el que se analizaron 50 muestras de sangre de cordón umbilical de gestantes atendidas en dos IPS, una de Bogotá y otra de Ubaté, (Cundinamarca), durante un período de 7 meses del año 2013. La selección de estas muestras fue basada en un muestreo no probabilístico. Se calculó el coeficiente de correlación de Pearson con su respectiva significancia estadística entre las variables cuantitativas de la madre y del neonato, y el volumen, el recuento de células CD34+ y el recuento total de células nucleadas por μL. Resultados: Se encontró correlación positiva entre la longitud del cordón y el recuento total de células nucleadas. Así mismo entre el volumen inicial de la muestra y el recuento total de células nucleadas y el recuento de células CD34+ en μL y en mL (mililitro). Discusión: Múltiples estudios se han desarrollado entorno a la búsqueda de variables maternas y neonatales que afectan la calidad de la muestra. En Colombia no se habían descrito las correlaciones encontradas en el presente estudio. Es por ello, que el desarrollo de nuevos estudios con diseños analíticos será enriquecedor.

Background: The cord blood has been recognized as a source of hematopoietic stem. Multiple studies have been conducted in order to determine maternal and neonatal variables that affect the volume, total nucleated cell count and CD34+ cells. This study correlated maternal and neonatal variables with the total nucleated cell count and CD34+ cells measured by μL. Materials and methods: Correlational study in which 50 samples of umbilical cord blood were analyzed of pregnant women at two health institutions, one at Bogotá and the other one at Ubaté, Cundinamarca, for a period of seven months of 2013. The selection of these samples was based on a non-probability sample. The Pearson correlation coefficient was calculated with their respective statistical significance between quantitative variables of mother and newborn, and the volume, the count of CD34+ cells and the total nucleated cell count measured by μL. Results: Positive correlation between the length of the umbilical cord and the total nucleated cell count was found. Similarly, correlation between total nucleated cell count with the initial volume and count of CD34+ cells in μL and mL was found. Discussion: Several studies have been conducted around to look maternal and neonatal variables that affect the sample quality, however, the Colombian literature has not described the correlation found in this study, so it requires the development of new research with designs of analytical studies to establish associations between the variables described.

Introdução: O sangue do cordão umbilical (SCU) tem sido reconhecido como uma fonte de células-tronco hematopoiéticas. Vários estudos têm sido realizados com a finalidade de determinar as variáveis maternas e neonatais que afetam o volume e a contagem total de células nucleadas e células CD34+. O presente estudo correlaciona as variáveis maternas e neonatais com a contagem total das células nucleadas e células CD34+ medidas em μL (microlitro). Materiais e métodos: Estudo correlacional no que 50 amostras de sangue do cordão umbilical de mulheres grávidas atendidas em dois IPs foram analisadas, uma em Bogota e outra em Ubaté (Cundinamarca), durante um período de 7 meses do ano 2013. A seleção destas amostras foi baseada em uma amostragem não probabilística. O coeficiente de correlação de Pearson foi calculado com a respectiva significância estatística entre variáveis quantitativas da mãe e do recém-nascido, assim como o volume, a contagem de células CD34 + e contagem total de células nucleadas por μL. Resultados: Correlação positiva foi encontrada entre o comprimento do cordão e a contagem total de células nucleadas. Também entre o volume da amostra inicial e contagem total de células nucleadas e contagem de células CD34+ em μL e em mL (mililitro). Discussão: Vários estudos têm sido desenvolvidos em torno da busca de variáveis maternas e neonatais que afetam a qualidade da amostra. Na Colômbia não tinham sido descritas as correlações encontradas no presente estudo. É por esta razão que será gratificante o desenvolvimento de novos estudos com desenhos analíticos.

The levels of pro-inflammatory factors are significantly decreased in cerebral palsy patients following an allogeneic umbilical cord blood cell transplant.

BACKGROUND AND OBJECTIVES: The transplantation of human umbilical cord blood cells (hUCBCs) has been shown to attenuate the unregulated activation of microglia in a rat model of cerebral palsy (CP). To investigate whether hUCBCs transplantation is also anti-inflammatory in humans, we performed a clinical trial in patients with CP. METHODS AND RESULTS: Allogeneic or autologous hUCBCs and erythropoietin (EPO) were intravenously injected into human patients with CP (mean age of approximately 38 weeks), and patients were analyzed for their motor function and social behavior. Blood samples were tested for cytokine levels. The most surprising finding in the study was that the cytokine levels were dependent on the donor cell source (allogeneic or autologous). Interestingly, the allogeneic treatment group demonstrated significantly decreased levels of pro-inflammatory factors, such as IL-1α, IL-6, TNF-ß, and RANTES, and showed a statistically significant improvement in motor and social behavior compared to the autologous treatment group. CONCLUSIONS: Given that inflammation plays a pivotal role in CP, our results suggest that allogeneic hUCBCs therapy may be an appropriate strategy for CP treatment. In addition, prior to transplantation, a detailed analysis of the amount of proinflammatory cytokines in cord blood may be needed to avoid exacerbating inflammatory responses.

The Levels of Pro-Inflammatory Factors Are Significantly Decreased in Cerebral Palsy Patients Following an Allogeneic Umbilical Cord Blood Cell Transplant

BACKGROUND AND OBJECTIVES: The transplantation of human umbilical cord blood cells (hUCBCs) has been shown to attenuate the unregulated activation of microglia in a rat model of cerebral palsy (CP). To investigate whether hUCBCs transplantation is also anti-inflammatory in humans, we performed a clinical trial in patients with CP. METHODS AND RESULTS: Allogeneic or autologous hUCBCs and erythropoietin (EPO) were intravenously injected into human patients with CP (mean age of approximately 38 weeks), and patients were analyzed for their motor function and social behavior. Blood samples were tested for cytokine levels. The most surprising finding in the study was that the cytokine levels were dependent on the donor cell source (allogeneic or autologous). Interestingly, the allogeneic treatment group demonstrated significantly decreased levels of pro-inflammatory factors, such as IL-1alpha, IL-6, TNF-beta, and RANTES, and showed a statistically significant improvement in motor and social behavior compared to the autologous treatment group. CONCLUSIONS: Given that inflammation plays a pivotal role in CP, our results suggest that allogeneic hUCBCs therapy may be an appropriate strategy for CP treatment. In addition, prior to transplantation, a detailed analysis of the amount of proinflammatory cytokines in cord blood may be needed to avoid exacerbating inflammatory responses.

Short & Long-term Repopulating Capacity of Cord Blood Cells That Are Stored Overnight before Cryopreservation / 대한수혈학회지

BACKGROUND: Umbilical cord blood (UCB) is generally stored overnight and it undergoes a CD34 positive selection process the next day for reducing the cost and due to the convenience. We intended to determine whether overnight storage of cord blood cells affects the short and long-term repopulating capacity. METHODS: Five individuals' UCB samples were analyzed by colony assay, apoptotic cell counts and long term bone marrow culture. All the samples were divided to four groups, which were the fresh group (immediate use of harvest), the overnight storage group (overnight storage at room temperature after harvest), the immediate cryopreservation group (immediate cryopreservation after harvest) and the overnight cryo group (cryopreservaton after overnight storage at room temperature after harvest). RESULTS: The number of colony forming units-granulocyte macrophage (CFU-GM) was 116.2+/-20.1 in the fresh group and 90.8+/-15.8 in the overnight storage group (P=0.07). The number of CFUs-GM was similar between the immediate and overnight cryo groups (P=0.79). The immediate cryo group showed a significantly lower number of CFUs-GM as compared to that of the fresh group (P=0.03). The apoptotic cells were detected at 21+/-6.8% in the fresh group and this was 24.2+/-2.4% in the overnight storage group (P=0.32), and this was similar between immediate and overnight cryo groups (P=0.80). The fresh group had a significantly lower number of apoptotic cells compared to that of the immediate cryo group (P=0.02). After long term stromal-based culture, the mean production of CFU-GM colonies was similar between all the groups (P>0.05). CONCLUSION: These results support the continue use of overnight storage of UCB before cryopreservation as a convenient, cost reducing measure.

Transplantation of Human Umbilical Cord Blood Improves Neurological Outcomes in the Rats after Traumatic Spinal Cord Injury

OBJECTIVE: Recent advances in stem cell biology make it possible to induce the regeneration of injured axons and to replace lost cells in the injured spinal cord. It has been found that stem cells in human cord blood differentiate into mature neurons and glial cells both in vitro and in vivo. These findings suggest that human umbilical cord blood cells(HUCBs) can be used as therapeutic donor cells in cases of spinal cord injury. METHODS: To attempt the repair an injured cord following spinal cord injury(SCI), we transplanted HUCBs into contused spinal cords. This was found to promote a long-term improvement in neurologic function relative to a lesion-control group. HUCBs were cultured in vitro for 7 days. Bromodeoxyuridine(BrdU) was added to the media to allow the BrdU to integrate into dividing cells. Cultured HUCBs(2x106 cells) were then injected into the injury epicenter 7 days after SCI. The Basso-Beattie-Bresnahan(BBB) locomotor rating system was used to score functional improvement in HUCBs transplanted rats. Immunohistochemical staining for neurofilament, macrotubule associated protein 2(MAP-2), glial fibrillary acidic protein(GFAP), and nestin was performed. RESULTS: Immunohistochemical analysis 5 weeks after SCI showed that gliogenesis of the transplanted donor HUCBs had occurred within the adult rat spinal cord. These donor-derived astrocyte-like cells extended their processes into the host tissues and integrated well. HUCBs derived neurons(neurofilament, MAP-2) and nestin expressing cells were also detected. Behavior analysis using BBB rating scores showed that functional improvement was greater in transplanted rats than in non-treated rats. CONCLUSION: HUCBs are one of the potential sources for transplantation material for the treatment of SCI.

The Effect of Cord Blood Plasma on Hematopoietic Colony Formation / 대한소아혈액종양학회지

PURPOSE: Umbilical cord blood transplantation is a alternative method as new hematopoietic stem cell transplantation and has been performed clinically in indicated disease. However, it have the problems for long-term storage of cord blood in liquid nitrogen and for limited application to adult due to small amount of hematopoietic stem cell. Therefore, several centers have carried out active research for ex vivo expansion of cord blood stem cell. We investigated the hematopoietic function of cord blood plasma for development of new techniques. METHODS: We acquired the nucleated cells of cord blood from healthy infant and bone marrow from healthy donor received granulocyte-colony stimulating factor. We evaluated hematopoietic colony formation according to source of stem cell and plasma by semisolid culture medium. Three experimental groups were divided as source of plasma: group for cord plasma, group for bone marrow plasma, group for mixture of cord plasma and bone marrow plasma. RESULTS: The results were as follows: 1) The colony formation according to source of stem cell in commercialized standard semisolid culture medium showed that cord blood in the number of CFU-GM was less than bone marrow, but not significantly different in CFU-GEMM. 2) The colony formation according to source of stem cell in semisolid culture medium using experimental plasma showed that cord blood in the number of CFU-GM was more than bone marrow. There were no cytotoxic effect of plasma to experimental cells. 3) The colony formation in semisolid culture medium contained plasma according to experimental group showed that the number of CFU-GM in cord blood plasma was significantly more than bone marrow plasma in spite of different source of stem cell. Conclusions: These results suggested that cord blood might contain enough hematopoiesis to enable to perform transplantation compared with bone marrow and, also, cord blood plasma might be contributed more effective colony formation than bone marrow plasma. Therefore, we propose that it may be good to store cord blood cells with cord blood plasma in long-term storage. We will investigate the composition of hematopoietic growth factors and cytokines in cord blood plasma and the effect of cord blood plasma for ex vivo expansion of cord blood cells.