ABSTRACT
RESUMEN Con el objetivo de evaluar criterios mínimos de celularidad de las unidades de sangre de cordón umbilical (USCU) según los estándares NETCORD en el Instituto Nacional Materno Perinatal de Lima, Perú, se realizó un estudio transversal que incluyó 100 USCU; se determinó el volumen, el recuento de células nucleadas totales (CNT) por hematología y el número de células CD34+ totales, así como también la viabilidad celular, por citometría de flujo. Se encontró que el 56% de las USCU no cumplen los umbrales mínimos de celularidad para ser criopreservadas en un banco de sangre de cordón umbilical. Se encontró, además, que las USCU de recién nacidos de mayor peso y de sexo femenino presentan mayor volumen y recuentos de células. En conclusión, es necesario considerar estas variables para optimizar la colecta de las USCU y obtener mayores recuentos de células que permita almacenar unidades de alta calidad en un futuro banco de sangre de cordón umbilical en Perú.
ABSTRACT A cross-sectional study that included 100 units of umbilical cord blood (UCB) was conducted to evaluate the minimum criteria of cellularity in UCB units, according to NetCord standards at Instituto Nacional Materno Perinatal in Lima, Peru. The volume, total count of nucleated cells by hematological tests and total number of CD34+ as well as cell viability by flow cytometry were determined. The study revealed that 56% of UCB units do not fulfill the minimum criteria of cellularity to be cryopreserved in an umbilical cord blood bank. Furthermore, the UCB units of newborns who weighed more and were female had a higher volume and cell count. In conclusion, these variables must undoubtedly be considered to optimize the collection of UCB units and obtain greater cell counts that enable the storage of high-quality units in a future umbilical cord blood bank in Peru.
Subject(s)
Female , Humans , Infant, Newborn , Pregnancy , Fetal Blood , Peru , Tissue Banks , Blood Cell Count , Cross-Sectional StudiesABSTRACT
UNLABELLED: : The pathophysiology of sepsis involves complex cytokine and inflammatory mediator networks. Downregulation of endothelial nitric oxide synthase contributes to sepsis-induced endothelial dysfunction. Human Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are known to reduce expression of proinflammatory cytokines and markers of apoptosis. We hypothesized that treatment with WJ-MSCs would protect renal, hepatic, and endothelial function in a cecal ligation and puncture (CLP) model of sepsis in rats. Rats were randomly divided into three groups: sham-operated rats; rats submitted to CLP and left untreated; and rats submitted to CLP and intraperitoneally injected, 6 hours later, with 1 × 10(6) WJ-MSCs. The glomerular filtration rate (GFR) was measured at 6 and 24 hours after CLP or sham surgery. All other studies were conducted at 24 hours after CLP or sham surgery. By 6 hours, GFR had decreased in the CLP rats. At 24 hours, Klotho renal expression significantly decreased. Treatment with WJ-MSCs improved the GFR; improved tubular function; decreased the CD68-positive cell count; decreased the fractional interstitial area; decreased expression of nuclear factor κB and of cytokines; increased expression of eNOS, vascular endothelial growth factor, and Klotho; attenuated renal apoptosis; ameliorated hepatic function; increased glycogen deposition in the liver; and improved survival. Sepsis-induced acute kidney injury is a state of Klotho deficiency, which WJ-MSCs can attenuate. Klotho protein expression was higher in WJ-MSCs than in human adipose-derived MSCs. Because WJ-MSCs preserve renal and hepatic function, they might play a protective role in sepsis. SIGNIFICANCE: Sepsis is the leading cause of death in intensive care units. Although many different treatments for sepsis have been tested, sepsis-related mortality rates remain high. It was hypothesized in this study that treatment with human Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) would protect renal, hepatic, and endothelial function in a model of sepsis in rats. Treatment with WJ-MSCs improved the glomerular filtration rate, improved tubular function, decreased expression of nuclear factor κB and of cytokines, increased expression of eNOS and of Klotho, attenuated renal apoptosis, and improved survival. Sepsis-induced acute kidney injury is a state of Klotho deficiency, which WJ-MSCs can attenuate.
Subject(s)
Acute Kidney Injury/prevention & control , Endothelium, Vascular/physiopathology , Liver Diseases/prevention & control , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Sepsis/surgery , Wharton Jelly/cytology , Acute Kidney Injury/etiology , Acute Kidney Injury/metabolism , Acute Kidney Injury/physiopathology , Animals , Apoptosis , Cells, Cultured , Cytokines/metabolism , Disease Models, Animal , Endothelium, Vascular/metabolism , Glomerular Filtration Rate , Glucuronidase/metabolism , Humans , Inflammation Mediators/metabolism , Kidney/metabolism , Kidney/pathology , Kidney/physiopathology , Klotho Proteins , Liver/metabolism , Liver/pathology , Liver/physiopathology , Liver Diseases/etiology , Liver Diseases/metabolism , Liver Diseases/physiopathology , Male , Mesenchymal Stem Cells/metabolism , NF-kappa B/metabolism , Phenotype , Rats, Wistar , Sepsis/metabolism , Sepsis/microbiology , Sepsis/physiopathology , Time FactorsABSTRACT
A cross-sectional study that included 100 units of umbilical cord blood (UCB) was conducted to evaluate the minimum criteria of cellularity in UCB units, according to NetCord standards at Instituto Nacional Materno Perinatal in Lima, Peru. The volume, total count of nucleated cells by hematological tests and total number of CD34+ as well as cell viability by flow cytometry were determined. The study revealed that 56% of UCB units do not fulfill the minimum criteria of cellularity to be cryopreserved in an umbilical cord blood bank. Furthermore, the UCB units of newborns who weighed more and were female had a higher volume and cell count. In conclusion, these variables must undoubtedly be considered to optimize the collection of UCB units and obtain greater cell counts that enable the storage of high-quality units in a future umbilical cord blood bank in Peru.
Subject(s)
Fetal Blood , Blood Cell Count , Cross-Sectional Studies , Female , Humans , Infant, Newborn , Peru , Pregnancy , Tissue BanksABSTRACT
In order to establish the reference intervals (RIs) of peripheral blood lymphocyte subsets (PBL) in healthy adults in Lima (Peru), a cross-sectional study was conducted among blood donors taken in between 2011 and 2012. Based on the criteria obtained from the guidelines of the Clinical and Laboratory Standards Institute (CLSI C28-A3), 318 samples were processed, 61.9% (197/318) coming from male donors. For PBL count, a flow cytometer with a simple platform was used. The RIs are established for each PBL in adults based on sex with their respective reference limits and 90% confidence intervals. Differences were found in CD3+ percentage counts (p=0.001) and in CD3-CD56+ absolute (p=0.003) and percentage counts (p?0.001). The RIs found are different to those described in studies conducted in other countries due to the characteristics of the population and the study model.
Subject(s)
Lymphocyte Subsets , Adolescent , Adult , Cross-Sectional Studies , Female , Humans , Lymphocyte Count , Male , Middle Aged , Peru , Reference Values , Urban Population , Young AdultABSTRACT
Con el objetivo de establecer los Intervalos de Referencia (InR) de las subpoblaciones linfocitarias (SPL) de sangre periférica en adultos sanos de Lima (Perú), se realizó un estudio transversal en donantes de sangre captados entre el 2011-2012. Siguiendo criterios basados en las directrices del Clinical and Laboratory Standards Institute (CLSI C28-A3), se procesaron 318 muestras, el 61,9% (197/318) de ellas procedentes de donantes de sexo masculino. Para el recuento de las SPL se empleó un citómetro de flujo de plataforma simple. Se establecieron los InR de cada SPL en personas adultas según sexo con sus respectivos límites de referencia y sus intervalos de confianza al 90%. Se encontraron diferencias en los recuentos porcentuales de CD3+ (p<0,01); y en los recuentos absolutos (p<0,01) y porcentuales (p=0,01) de CD3-CD56+. Los InR hallados son diferentes a los descritos en estudios realizados en otros países. Dichas diferencias se deben a las características de la población y al modelo de estudio.
In order to establish the reference intervals (RIs) of peripheral blood lymphocyte subsets (PBL) in healthy adults in Lima (Peru), a cross-sectional study was conducted among blood donors taken in between 2011 and 2012. Based on the criteria obtained from the guidelines of the Clinical and Laboratory Standards Institute (CLSI C28-A3), 318 samples were processed, 61.9% (197/318) coming from male donors. For PBL count, a flow cytometer with a simple platform was used. The RIs are established for each PBL in adults based on sex with their respective reference limits and 90% confidence intervals. Differences were found in CD3+ percentage counts (p=0.001) and in CD3-CD56+ absolute (p=0.003) and percentage counts (p?0.001). The RIs found are different to those described in studies conducted in other countries due to the characteristics of the population and the study model.
