Human platelet lysate (PL) and human platelet lysate serum (PLS) are alternatives to fetal bovine serum (FBS) due to their ethical concerns, variability between batches, and their possible introduction of xenogenic contaminants. This study compared the composition and efficacy of PL, PLS and FBS as supplements in the culture and cryopreservation of human dermal fibroblasts, Wharton’s jelly derived mesenchymal stem cells (WJ-MCS), and adipose tissue (AdMSC). Biochemical components, some growth factors and cytokines present in each of them were analyzed, in addition, the cells were cultured in media supplemented with 5% PL, 5% PLS and 10% FBS, and exposed to different freezing and thawing solutions with the supplements under study. Biochemical parameters were found to be similar in PL and PLS compared to FBS, with some differences in fibrinogen and calcium concentration. Growth factors and cytokines were higher in PL and PLS compared to FBS. Cell proliferation and morphology showed no significant differences between the three-culture media. Regarding the cryopreservation and thawing of cells, better results were obtained with PLS and FBS. In conclusion, PL and PLS are an excellent choice to replace the standard supplement of animal origin (FBS) in the media used for the culture and cryopreservation of fibroblasts, WJ-MSC and AdMSC.
Se realizó un estudio cuasi experimental; se recolectaron 25 cordones umbilicales (UC) en solución de transporte, de los cuales sólo nueve cumplieron con los criterios de inclusión. El tamaño de los cordones fue mayor de 30 cm con un diámetro de ≥1.5 cm; éstos fueron seccionados en 15 fragmentos de aproximadamente 2 cm, y expuestos a diferentes temperaturas (10°C, 22°C y 32°C) durante 6, 12, 24, 48 y 72 horas. Se obtuvo una cantidad de 600 mg de gelatina de Wharton (WJ) de cada fragmento. Se analizó la celularidad y viabilidad con la tinción de azul de tripán. Los fragmentos mantenidos a 10°C presentaron una media de viabilidad de 85,6% ± 6,8 después de 6 horas, con una pérdida de viabilidad a las 72 horas, de 13,6% (p=0,001); a 22ºC la media de viabilidad fue de 84,3% ± 4,2 con una pérdida de viabilidad a las 72 horas de 27.4% (p<0,001); a 32ºC 83,6 % ± 4,6 con una disminución del 60,4% a las 72 horas (p<0,001). Respecto al número de células/mg de WJ, a 22ºC se presentó una media de 167 cel./mg ± 36,5 a las 6 horas, con una pérdida de células a las 72 horas de 112 cel./mg (p=0,001); a 32°C 139 cel./mg ± 73,7, con una pérdida de 98 cel./mg a las 72 horas (p=0,003). La celularidad no fue estadísticamente significativa a 10°C entre los diferentes períodos (p=0,931). Los datos observados en este estudio indican que la temperatura ideal para la preservación y transporte del tejido de UC es de 10°C, con un tiempo máximo de conservación de 24 horas. Pre-processing temperature and time: essential factors for cellular performance from Wharton's gelatin Abstract A quasi-experimental study was carried out, where 25 umbilical cords (UC) were collected in transport solution of which only nine (9) fulfilled the inclusion criteria. The size of the cords was greater than 30 cm with a diameter of ≥ 1.5 cm; these were cut into 15 fragments of approximately 2 cm and exposed to different temperatures (10 ° C, 22 ° C and 32 ° C) for 6, 12, 24, 48 and 72 hours. It was obtained 600 mgs of Wharton's gelatin (WJ) from each fragment. Cellularity and viability were analyzed with trypan blue staining. The fragments maintained at 10 ° C presented an average viability of 85.6% ± 6.8 at 6 hours, with a loss of viability at 72 hours of 13.6% (p = 0.001). At 22 ° C the viability average was 84.3% ± 4.2 with a loss of viability at 72 hours of 27.4% (p <0.001). At 32ºC 83.6% ± 4.6 with a decrease of 60.4% at 72 hours (p <0.001). With respect to the number of cells / mg of WJ, at 22 ° C an average of 167 cells / mg ± 36.5 was presented at 6 hours with a cell loss at 72 hours of 112 cells / mg (p = 0.001). ). At 32 ° C 139 cel./mg ± 73.7, with a loss of 98 cel./mg at 72 hours (p = 0.003). The cellularity was not statistically significant at 10 ° C between the different times (p = 0.931). The data found in this study indicate that the ideal temperature for the preservation and transport of UC tissue is 10 ° C, with a maximum conservation time of 24 hours.
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