Molecules of animal or bacterial origin, which pose a risk for zoonoses or immune rejection, are commonly used for extraction, culture, and cryopreservation of mesenchymal stem cells. There is no sequential and orderly protocol for producing human adipose-derived stem cells (hASCs) under xeno-free conditions. After standardizing a human platelet lysate (hPL) production protocol, four human adipose tissue samples were processed through explants with fetal bovine serum (FBS)-supplemented or hPLsupplemented media for extracting the adipose-derived stem cells. The cells were cultivated in cell culture medium + hPL (5%) or FBS (10%). The cellular replication rate, immunophenotype, and differentiation potential were evaluated at fourth passage. Cellular viability was evaluated before and after cryopreservation of the cells, with an hPL-based solution compared with an FBS-based solution. The explants cultured in hPL-supplemented media showed earlier and faster hASC proliferation than did those supplemented with FBS. Likewise, cells grown in hPL-supplemented media showed a greater proliferation rate, without losing the immunophenotype. Osteogenic differentiation of xeno-free hASC was higher than the hASC produced in standard conditions. However, adipogenic differentiation was reduced in xeno-free hASC. Finally, the cells cryopreserved in an hPL-based solution showed a higher cellular viability than the cells cryopreserved in an FBS-based. In conclusion, we have developed a complete xeno-free protocol for extracting, culturing, and cryopreserving hASCs that can be safely implemented in clinical studies. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:358-365 SIGNIFICANCEThis study was performed to standardize a complete ordered protocol to produce xeno-free human adipose-derived mesenchymal stem cells (hASCs) as a safe therapeutic alternative. Cells were extracted by adipose tissue explants and then cultured and cryopreserved using human platelet lysate (hPL). Different scientific journals have published data regarding the use of hPL as a safe fetal bovine serum substitute for hASC culture, using heparin to avoid clot formation. This article reports the use of hPL for extracting, culturing, and cryopreserving hASCs without anticoagulant.

Introduction: Major depression (MD), like other mood disorders, is considered a worldwide endemic pathology, becoming one of the biggest public health problems. The hereditary factors of mood disorders have been studied for many years and different chromosomal regions and genes have been involved in this physiopathological process. Objective: To establish the association between allelic variants of the serotonin transporter gene (5-HTTLPR and VNTR) and MD in a population with this diagnostic in the department of Caldas, Colombia. Materials and methods: A case-control study was conducted with individuals older than 16 years of age born in the department of Caldas. The sample was composed of 59 patients with the MD with family antecedents of the pathology and 59 controls paired by precedence, age, and gender. For the cases and controls selection the Diagnostic Interview for Genetic Studies (DIGS) was used. Using the Hranilovic et al. protocol, the polymorphic regions in the promoter and third intron of the Serotonin Transporter gene was amplified. Results: It was not possible to find association between MD and the genetic or clinical variables. The absence of the short allele of the promoter could act as a protective factor (OR=0.70 CI 95%=0.313 to 1.604), for the development of the pathology in this population, and the presence of at least one copy of the 10 repetition alleles of the third intron could act as a risk factor (OR: 1.25), but the wide confidence interval (CI 95%=0.38 to 2.64) does not permit supporting these conclusions. Discussion: The results obtained in this population do not yield conclusive information related with the etiopathogeny of MD, but do not contradict those obtained in other studies with bigger samples than ours. The broad confidence interval does not support conclusions about the role as a risk factor for the 10 repetition alleles of the intron or the absence of the S allele of the promoter as a protector factor. Further studies with larger population samples may help to clarify these facts.

Candida albicans is fungus capable of changing from yeast to filamentous form when it’s transformed from a normal commensal to an opportunistic pathogen. The development of alternatives that interfere with this transition could be an effective way to reduce candidiasis. In this regard, evaluate the inhibitory effect of two Borojoa patinoi silver nanoparticles (AgNPs) produced by green synthesis at 5 °C and 25 °C on the process of filamentation of Candida albicans. The percentage of inhibition of filamentous forms of C. albicans ATCC10231 and C. albicans SC5314 with AgNPs was determined. Results showed that temperature of synthesis affected both the shape and size of silver nanoparticles synthesized using Borojoa patinoi extracts. The inhibition percentage of filamentous forms of Candida albicans ATCC10231 when treated with silver nanoparticles synthesized at 5 °C was 85.9% and at 25 °C it was 40%. C. albicans SC5314 when treated with AgNP synthesized at 5 °C was 97.2% and at 25 °C it was 64%. Cell toxicity assay showed that at 100ng/ml, AgNPs synthesized at 25 °C were safe in MES-OV CRL-3272 cell line. Our results showed that the silver nanoparticles obtained from Borojoa patinoi are inhibitors of the filamentous process of C. albicans.

Desde su descripción inicial, hace ya más de 40 años, las células madre mesenquimales (MSC) fueron reconocidas como una importante alternativa para el manejo de enfermedades caracterizadas por la pérdida aguda o crónica de tejido, gracias a su capacidad de proliferación y diferenciación, lo cual les permitiría sustituir las células perdidas y de esta forma recuperar la estructura y función. Cada vez es más abundante la evidencia que sugiere el potencial de estas células para el manejo de un amplio grupo de enfermedades, al menos en modelos experimentales preclínicos. No obstante, esta capacidad no ha podido refrendarse contundente y consistentemente en ensayos clínicos. Con la presente revisión, se pretende presentar una visión del estado actual del desarrollo conceptual en torno a las capacidades terapéuticas de las MSC y un análisis crítico de algunos de los factores, que han impedido que estas sean una opción terapéutica usable en la práctica clínica diaria.