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-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.