RESUMEN
The use of human mesenchymal stem cells (hMSC) in clinical applications has been increasing over the last decade. However, to be applied in a clinical setting hMSC need to comply with specific requirements in terms of identity, potency and purity. This study reports the improvement of established tangential flow filtration (TFF)-based washing strategies, further increasing hMSC purity, using negative mode expanded bed adsorption (EBA) chromatography with a new multimodal prototype matrix based on core-shell bead technology. The matrix was characterized and a stable, expanded bed could be obtained using standard equipment adapted from what is used for conventional packed bed chromatography processes. The effect of different expansion rates on cell recovery yield and protein removal capacity was assessed. The best trade-off between cell recovery (89%) and protein clearance (67%) was achieved using an intermediate expansion bed rate (1.4). Furthermore, we also showed that EBA chromatography can be efficiently integrated on the already established process for the downstream processing (DSP) of hMSC, where it improved the washing efficiency more than 10-fold, recovering approximately 70% of cells after global processing. This strategy showed not to impact cell viability (>95%), neither hMSC's characteristics in terms of morphology, immunophenotype, proliferation, adhesion capacity and multipotent differentiation potential.
Asunto(s)
Cromatografía , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/citología , Supervivencia Celular , Filtración , HumanosRESUMEN
NMR cryoporometry is a unique method permitting the investigation of pores in the microporous and mesoporous regimes for samples in aqueous environments. Here, we apply the technique to porous biodegradable polymer microparticles designed as devices for drug delivery in depot formulations. The results indicate that structural features too small to be captured in surface and fracture images obtained by SEM are able to be accessed using the technique, and that the evolution of pore structure can be studied for several days as the particles swell and degrade in the aqueous environment.
Asunto(s)
Materiales Biocompatibles/química , Ácido Láctico/química , Espectroscopía de Resonancia Magnética/métodos , Ácido Poliglicólico/química , Polímeros/química , Porosidad , Química Farmacéutica/métodos , Portadores de Fármacos , Microscopía Electrónica de Rastreo , Tamaño de la Partícula , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Propiedades de Superficie , Factores de Tiempo , Agua/químicaRESUMEN
Stress development during drying of coatings produced from aqueous dispersions of calcium carbonate particles in the presence and absence of organic binders was studied using a controlled-environment stress apparatus that simultaneously monitored drying stress, weight loss, and relative humidity. Specifically, the influence of two organic binders on drying stress evolution was investigated: (1) carboxymethylcellulose, a water-soluble viscosifying aid, and (2) a styrene-butadiene latex emulsion of varying glass transition temperature. The stress histories exhibited three distinct regions. First, a period of stress rise was observed, which reflected the capillary tension exerted by the liquid on the particle network. Second, a maximum stress was observed. Third, it was followed by a period of either stress decay or rise depending on the organic species present. Significant differences in stress histories were observed between coatings containing soluble and nonsoluble binders. Maximum drying stresses (sigmamax) of 0.2-0.5 MPa were observed for coatings produced from pure calcium carbonate or calcium carbonate-latex suspensions, whereas coatings with carboxymethylcellulose exhibited substantially higher sigmamax values of 1-2 MPa. Upon drying, these coatings were quite hygroscopic, such that cyclic variations in relative humidity induced large cyclic changes in residual stress.