Long-Term Quality Control Program Plan for Cord Blood Banks in Korea: A Pilot Study for Cryopreservation Stability

BACKGROUND: Maintaining the quality of cryopreserved cord blood is crucial. In this pilot study, we describe the results of the internal quality control program for a cord blood bank thus far. METHODS: Donated cord blood units unsuitable for transplantation were selected for internal quality control once a month. One unit of cord blood, aliquoted into 21 capillaries, was cryopreserved and thawed annually to analyze the total nucleated cell count, CD34⁺ cell count, cell viability test, and colony-forming units assay. RESULTS: No significant differences in the variables (total nucleated cell count, cell viability, CD34⁺ cell count) were observed between samples cryopreserved for one and two years. Upon comparing the variables before cryopreservation and post thawing with the capillaries of one year of storage, cell viability and CD34⁺ cell counts decreased significantly. The use of cord blood samples in capillaries, which can be easily stored for a long period, was similar to the methods used for testing segments attached to the cord blood unit. CONCLUSIONS: The results of this study may be useful for determining the period during which the quality of cryopreserved cord blood units used for transplantation is maintained.

Rapid thawing human sperm does not affect basic parameters in normozoospermic men: a double-blind prospective study

PURPOSE: To compare sperm recovery from slow versus rapid thawing technique using thirty-eight normozoospermic human sperm samples, as follows. Twenty-one samples from men taking part in routine infertility screening exams (infertile group) and seventeen from proven fertile volunteer men with at least one child (fertile group). MATERIALS AND METHODS: After analysis of motility, concentration, strict morphology and functional integrity of membranes, sperm was divided into two aliquots of 0.5 mL each and frozen in TyB-G medium. Samples were thawed at room temperature (25 ± 2º C) for 25 minutes (slow thaw) or in a water bath at 75º C for 20 seconds followed by water bath at 37º C for 3 minutes (rapid thaw). After thawing, motility, strict morphology and functional integrity of membranes were evaluated by a blinded investigator. The results were expressed as mean ± standard deviation for parametric variables and analyzed using Student's t-test. Data with unpaired non-parametric variables were expressed as median (interquartile range) and analyzed by the Mann-Whitney test. Wilcoxon test was used to analyze non-parametric paired variables. RESULTS: There was no significant difference between techniques for total and progressive motility, percentage of normal morphological forms, hypoosmotic swelling test. CONCLUSIONS: Although the rapid thawing protocol was completed in a shorter time (three minutes and 20 seconds versus 25 minutes, respectively), it wasn't harmful since both techniques showed comparable spermatozoa recovery. Additional research is needed to confirm its safety in clinical research before introducing this methodology in routine assisted reproduction.

Evaluación de la contaminación microbiológica en productos de células progenitoras hematopoyéticas. Experiencia de un laboratorio de procesamiento de CPH y Banco de Sangre de Cordón Umbilical / Microbiological evaluation in hematopoietic progenitor cell products. Experience from an HPC processing laboratory and Cord Blood Bank

Los productos de células progenitoras hematopoyéticas (CPH) representan una fuente potencial de infección en pacientes inmunosuprimidos que reciben infusión de CPH como parte de su tratamiento. La probabilidad de contaminación de cada producto difiere según la técnica de colecta y el procesamiento aplicado. En este trabajo hemos realizado un análisis retrospectivo de los resultados de cultivos microbiológicos de 1707 productos de CPH obtenidos de sus tres fuentes (médula ósea, sangre periférica y sangre de cordón umbilical) con el objetivo de determinar la proporción de unidades contaminadas. Además, fueron comparadas las distintas técnicas de colecta y las diferentes manipulaciones a las que fueron sometidos los productos. Por otro lado, se analizaron las posibles fuentes de contaminación según el microorganismo identificado y se evaluó la supervivencia de los mismos luego del descongelamiento. La prevalencia de productos de CPH con cultivos microbiológicos positivos reportados en este estudio (5,2%) se corresponde con lo descripto en la literatura. No encontramos diferencias significativas al comparar los productos según la fuente de la cual provenían las CPH. Tampoco hubo diferencias según los procedimientos aplicados a cada unidad. Los microorganismos aislados en los productos de CPH fueron los esperados de acuerdo a la fuente de la cual provenían las células. Pudo comprobarse que algunos de ellos son capaces de sobrevivir a los procesos de criopreservación y descongelamiento. La estricta adhesión a las normas de buenas prácticas de manufactura y buenas prácticas tisulares es un requisito para minimizar los riesgos de introducir microorganismos contaminantes. Disponer de un producto de CPH seguro es fundamental para el éxito de un trasplante.

Hematopoietic stem cell products (HSCP) represent a potential source of infection for immunosuprressed patients that receive HSCP infusion as part of their treatment. The probability of contamination of a HSCP product depends on the collection technique as well as the processing performed. In this work we have carried out a retrospective analysis of the results of 1707 microbiological cultures of HSCP products obtained from three different sources: bone marrow, peripheral blood and umbilical cord blood. We determined the proportion of HSCP units that were contaminated by microorganisms. Furthermore we compared the results obtained with different collection techniques and with the distinct manipulations that were used for processing. Moreover we analysed the possible sources of contamination related to the microorganisms identified and we evaluated the survival of them after thawing. The proportion of HSCP products with positive microbiological cultures obtained in this study (5,2%) correlates with that reported by other authors. We have not found significant differences between the results achieved with HSCP products from different sources. There were neither differences depending on the procedures applied. The isolated microorganisms from the HSCP products were the expected in accordance with the source of the cells. It could be demonstrated that some of them were capable of surviving the cryopreservation and thawing processes. Adherence to good manufacture practices and good tissue practices regulations is critical for minimizing the risks of introducing contaminant microorganisms. A safe HSCP product is essential for the success of a transplant.

Controle microbiológico em valvas cardíacas humanas / Microbiologyc control in human heart valves

OBJETIVO: Avaliar, sob o aspecto microbiológico, valvas processadas pelo Banco de Valvas Cardíacas Humanas da Irmandade da Santa Casa de Misericórdia de Curitiba, para serem utilizadas em cirurgias cardiovasculares. MÉTODOS: Foi avaliado o processamento de 1.671 valvas, no período de junho de 1999 a junho de 2004. Das valvas e soluções envolvidas no processo foram coletadas amostras e semeadas nos meios de cultura: meio líquido tioglicolato, caldo soja tripticaseína e caldo Sabouraud, com quatorze dias de incubação, utilizando a metodologia modificada baseada na Farmacopéia Brasileira 1998 e USP 1990 (United States Pharmacopeia). Nas amostras que apresentaram crescimento foram realizadas as identificações microbianas. RESULTADOS: Em um total de 1.671 amostras analisadas, 92 por cento foram consideradas próprias para utilização, sob o aspecto microbiológico, uma vez que não apresentaram contaminação microbiana. Somente 8 por cento não foram liberadas para uso clínico por motivo de contaminação em alguma etapa do processamento da valva. CONCLUSÃO: Analisando os resultados, observou-se a importância do controle microbiológico em enxertos humanos, evitando a utilização de valvas com contaminação microbiológica em pacientes submetidos à cirurgia cardiovascular.

OBJECTIVE: To evaluate, from microbiological point of view, the valves processed by Human Heart Valve Bank of Santa Casa de Misericórdia of Curitiba for use in cardiovascular surgeries. METHODS: The processing of 1,671 valves, accomplished within the period of time between July 1999 and June 2004, was evaluated. Out of the valves and the solutions involved in the process, samples were collected and spread in culture mediums, such as fluid thioglycollate medium, tryptic soy broth and Sabouraud broth, for incubation during 14 days, using a modified methodology based on the Farmacopéia Brasileira 1988 (Brazilian Pharmacopeia) and USP 1990 (United States Pharmacopeia). The samples in which growing was observed were submitted to microbian identification. RESULTS: In a set of 1,671 samples, 92 percent were considered proper for use under microbiological point of view, since they did not display microbian contamination. The remaining 8 percent were rejected for clinical use because of contamination in some stage of the valve processing. CONCLUSION: From the Analysis of the results, it was observed the importance of microbiological control in human grafts, in order to avoid using microbiologically contaminated valves in patients submitted to cardiovascular surgery.

Comparative study of cryopreserved bone tissue and tissue preserved in a 98 percent glycerol solution / Estudo comparativo entre o tecido ósseo criopreservado e o conservado em glicerol a 98 por cento

OBJECTIVE: To compare the bone graft cryopreservation method (at -80°C) with a preservation method using a 98 percent glycerol solution at room temperature (10°C-35°C), by testing the antibacterial and fungal effects of 98 percent glycerol and comparatively analyzing the observed histological changes resulting from the use of both methods. METHOD: This study was of 30 samples of trabecular bone tissue from 10 patients undergoing total hip arthroplasty. Each femoral head provided 3 samples that were randomized into 3 groups, namely, the control group, the cryopreserved group, and the group preserved in a 98 percent glycerol at room temperature for 1 year. The samples were submitted to histomorphologic, cell feasibility, and microbiologic analyses. The results were statistically analyzed using the McNemar test, with a statistical significance index of 0.05. RESULTS: Values obtained using the McNemar test to compare probability distributions of histomorphologic variables (mature or lamellar bone, immature bone, and necrosis) and cell feasibility (osteoblasts and osteoclasts) indicated that there is no difference between the distributions of variables under the 3 experimental conditions. Microbiological analysis of the 98 percent glycerol solution and bone fragments from samples stored for 1 year at room temperature did not show bacterial or fungal growth. The histological and microbiological investigation were performed at 2 different time points: immediately after the sample processing and after 1 year. CONCLUSION: The method used to preserve bone grafts kept in 98 percent glycerol at room temperature (10°C-35°C) was similar to cryopreservation in terms of bone matrix preservation; no bacteria or fungi were found in the samples.

OBJETIVO: Comparar o método da criopreservação de enxertos ósseos (- 80° C) com o da conservação em glicerol a 98 por cento em temperatura ambiente (10° C a 35° C), testando os efeitos antibacterianos e antifúngicos do glicerol a 98 por cento e analisando comparativamente as alterações histológicas verificadas e decorrentes do emprego dos dois métodos. MÉTODO: Este estudo foi constituído de 30 amostras de tecido ósseo trabecular provenientes de 10 pacientes, submetidos a Artroplastia Total do Quadril. Cada cabeça femoral forneceu 3 amostras e estas foram divididas aleatoriamente em 3 grupos, a saber: controle, criopreservado e conservado em glicerol a 98 por cento à temperatura ambiente durante um ano. As amostras foram encaminhadas à Anatomia Patológica para estudo histomorfologico, de viabilidade celular, e microbiológico. Os resultados foram analisados estatisticamente pelo método de McNemar, com índice de significância de 0,05. RESULTADOS: A análise dos valores obtidos no teste de McNemar na comparação das distribuições de probabilidades das variáveis da histomorfologia (osso maduro ou lamelar, osso imaturo e necrose) e da viabilidade celular (osteoblastos e osteoclastos) indica não haver diferença entre as distribuições das variáveis nas três condições experimentais. A análise microbiológica da solução de glicerol a 98 por cento e dos fragmentos ósseos das amostras armazenadas durante um ano em temperatura ambiente não apresentou crescimento bacteriano ou de fungos. As espécimens do grupo controle foram analisadas histológica e microbiologicamente logo após a coleta das mesmas. CONCLUSÃO: O método de conservação de enxertos ósseos mantidos no glicerol a 98 por cento em temperatura ambiente (10°C a 35°C) foi similar ao da criopreservação quanto à preservação da matriz óssea e à ausência de crescimento de bactérias ou fungos.

Preservación de eritrocitos y cambios físicos ocurridos durante el almacenamiento / Preservation of erythrocytes and happened physical changes during storage

La preservación de sangre y de sus derivados ha sido una constante en el desarrollo de los bancos de sangre modernos. Los dos métodos actualmente utilizados son la fase líquida y la criopreservación. Se revisaron las generalidades de cada uno de ellos, así como los cambios físicos ocurridos a los eritrocitos durante su almacenamiento

Banco de tecidos: estruturação e normatização / Tissue bank: structure and organization

Os autores descrevem os passos que foram necessários à reestruturação do Banco de Tecidos do IOT do Hospital das Clínicas da Faculdade de Medicina da USP, realizados na instituição, onde prestam serviços. Citam a lei nº 9.434 de 4 de fevereiro de 1997, como diretriz de todo o processo de mudança e na confecção dos termos de doação e recepção, fundamentais no exercício das atividades de um banco de tecidos. Descrevem também a nova estrutura física e a de recursos humanos, utilizados nas atividades de captação, processamento e crio-preservação. É enfatizado que a atuação ritmada da equipe é que permitirá melhor qualidade dos aloenxertos e, conseqüentemente, assistência mais efetiva ao paciente.

Ley de reproducción asistida: opciones para su formulación / Assisted reproduction law: options for it formulation

Las variables técnicas que puede tener un aley de reproducción asistida son, como puede apreciarse, múltiples. Determinar una u otra opción, importa, básicamente, juicios de valor, cuyos fundamentos últimos no son criterios biológicos, sino de otros ámbitos, filosóficos, morales, religiosos, sociológicos. No puede concebirse, entonces, una legislación dictada sólo bajo criterios médicos, pero no es menos cierto, que debe contarse con la información biológica básica que permita fundamentar adecuadamentedichos juicios de valor. este artículo pretende ser un aporte en ese sentido

Effect of plasma freezing temperature, anticoagulant and time of storage on factor VIII: C activity in cryoprecipitate

The influence of ACD and CPDA-1 anticoagulants, and storage time for 3 and 6 months on F VIII:C activity were compared in cryprecipitate obtained at -70ºC, and -30ºC plasma freezing temperature. To eliminate variations in F VIII:C activity between donor plasma, the cryoprecipitation at -70ºC and -30ºC was made in paired plasma volumes (approximately 100 ml) from each blood unit. Employing ACD plasmas (n= 50), there was no significant difference in F VIII:C activity between cryoprecipitate prepared at -70ºC (X=31.1 IU/bag) and -30ºC was made in paired plasma volumes (approximately 100 ml) from each blood unit. Employing ACD plasmas (n= 50), there was no significant difference in F VIII:C activity between cryprecipitate prepared at -70ºC (X= 31.1 IU/bag) and -30ºC (X = 30.5 IU/bag), and the storage did not modifify FVIII:C activity. In contrast, in cryprecipitate prepared from CPDA-1 plasma (n= 31), the F VIII:C levels obtained at-30ºC (X= 43.8IU/bag) were significantly higher than those at-70ºC (X=37.3 IU/bag), but a deterioration of F VIII:C activity (about 50 percent) was observed after 6 months of cryprecipitate storage. Therefore, if cryprecipitate is stored it stored it would be more convenient to use ACD instead of CPDA-1 and make cryoprecipitation either at-70ºC or-30ºC

Aloinjertos osteocondrales: comparacion de diferentes metodos de preservacion en un modelo animal / Osteochondral allogfrafts: comparison of different methosd of preservation in an animal model

Se comparan diferentes metodos de preservacion apolicados a cartilago articular proveniente de conejos Nueva Zelandia: criopreservacion en Dimetilsulfoxido al 10 por ciento por 6 horas y congelacion lenta hasta -76oC, congelacion sin criopreservacion y refrigeracion a 4 oC encontrando que la criopreservacion y congelacion lenta ofrece mejores resultados con una viabilidad celular del 25 por ciento luego de una semana de almacenamiento. En una segunda base "in vitro" se tomaron 3 grupos de conejos (6 en cada grupo) que recibieron un aloinjerto osteocondrial del condilo femoral externo. En el primer grupo se usaron injertos frescos, en el segundo injertos refrigerados a 4oC por una semana y en el tercero injertos criopreservados en dimetilsulfoxido y congelados lentamente y posteriormente almacenados a -76oC por una semana. Los resultados funcionales a corto plazo son mejores en el tercer grupo. Se espera informar los resultados funcionales y morfologicos luego de un periodo de seguimiento de un ano