Good manufacturing practice-grade production of unrestricted somatic stem cell from fresh cord blood.

BACKGROUND AIMS: The discovery of unrestricted somatic stem cells (USSC), a non-hematopoietic stem cell population, brought cord blood (CB) to the attention of regenerative medicine for defining more protocols for non-hematopoietic indications. We demonstrate that a reliable and reproducible method for good manufacturing practice (GMP)-conforming generation of USSC is possible that fulfils safety requirements as well as criteria for clinical applications, such as adherence of strict regulations on cell isolation and expansion. METHODS: In order to maintain GMP conformity, the automated cell processing system Sepax (Biosafe) was implemented for mononucleated cell (MNC) separation from fresh CB. After USSC generation, clinical-scale expansion was achieved by multi-layered CellSTACKs (Costar/Corning). Infectious disease markers, pyrogen and endotoxin levels, immunophenotype, potency, genetic stability and sterility of the cell product were evaluated. RESULTS: The MNC isolation and cell cultivation methods used led to safe and reproducible GMP-conforming USSC production while maintaining somatic stem cell character. CONCLUSIONS: Together with implemented in-process controls guaranteeing contamination-free products with adult stem cell character, USSC produced as suggested here may serve as a universal allogeneic stem cell source for future cell treatment and clinical settings.

Stem cells derived from cord blood in transplantation and regenerative medicine.

BACKGROUND: Physicians of any specialty may be the first persons to whom prospective parents turn for information about the acquisition and storage of stem cells derived from cord blood. Stem cells can potentially be used to treat many diseases, yet they are not a panacea. This article provides an overview of their current and possible future applications. METHODS: Original papers were retrieved by a selective search of the literature, and the Internet sites and advertising brochures of private stem cell banks were also examined. RESULTS: Allogeneic hematopoietic stem cells derived from umbilical cord blood (obtained from healthy donors, rather than from the patient to be treated) have been in routine use worldwide for more than ten years in the treatment of hematopoietic diseases. Experiments in cell culture and in animal models suggest that these cells might be of therapeutic use in regenerative medicine, but also show that this potential can be realized only if the cells are not cryopreserved. There is as yet no routine clinical application of autologous hematopoietic stem cells from cord blood (self-donation of blood), even though cord blood has been stored in private banks for more than ten years. CONCLUSIONS: Autologous stem cells from cord blood have poor prospects for use in regenerative medicine, because they have to be cryopreserved until use. Physicians should tell prospective parents that they have no reason to feel guilty if they choose not to store cord blood in a private bank.

The SCINEXA: a novel, validated score to simultaneously assess and differentiate between intrinsic and extrinsic skin ageing.

BACKGROUND: Studies on the pathogenesis of skin ageing as well as efficacy testing of cosmetic and aesthetic measures to prevent or reverse skin ageing require - as an easy to use method - a validated non-invasive clinical score, which allows to simultaneously assess and differentiate between intrinsic (=chronological) and extrinsic (=photo-) skin ageing. Such an ideal score, however, does currently not exist. OBJECTIVES: We developed a novel skin ageing score 'SCINEXA' comprising 5 items indicative of intrinsic and 18 items highly characteristic of extrinsic skin ageing. These items were used to define an index (index(discr)) that allowed differentiating between intrinsic versus extrinsic skin ageing. In order to validate the 'SCINEXA', we asked whether it can be used to discriminate regular sunbed users, which have been chronically exposed to ultraviolet radiation and thus are prone to photoageing, from non-sunbed users, which were considered paradigmatic for intrinsic skin ageing. METHODS: For this purpose, 58 non-sunbed users and 16 regularly sunbed users were assessed. In addition to the clinical examination of the 23 score items potential confounders were considered by questionnaire. RESULTS: By employing the index(discr), we were able to classify 92% of all study subjects correctly as sunbed or non-sunbed users. Specifically, an index above 2 was associated with sunbed use and thus extrinsic skin ageing, whereas an index below 2 indicated intrinsic skin ageing. CONCLUSION: The novel 'SCINEXA' is suitable for the simultaneous assessment of intrinsic and extrinsic skin ageing.

Contribution of smoking and air pollution exposure in urban areas to social differences in respiratory health.

BACKGROUND: Socio-economic status, smoking, and exposure to increased levels of environmental air pollution are associated with adverse effects on respiratory health. We assessed the contribution of occupational exposures, smoking and outdoor air pollution as competing factors for the association between socio-economic status and respiratory health indicators in a cohort of women from the Ruhr area aged 55 at the time of investigation between 1985 and 1990. METHODS: Data of 1251 women with spirometry and complete questionnaire information about respiratory diseases, smoking and potential confounders were used in the analyses. Exposure to large-scale air pollution was assessed with data from monitoring stations. Exposure to small-scale air pollution was assessed as traffic-related exposure by distance to the nearest major road. Socio-economic status was defined by educational level. Multiple regression models were used to estimate the contribution of occupational exposures, smoking and outdoor air pollution to social differences in respiratory health. RESULTS: Women with less than 10 years of school education in comparison to more than 10 years of school education were more often occupationally exposed (16.4% vs. 10.1%), smoked more often (20.3% vs. 13.9%), and lived more often close to major roads (26.0% vs. 22.9%). Long-term exposure to increased levels of PM10 was significantly associated with lower school education. Women with low school education were more likely to suffer from respiratory symptoms and had reduced lung function. In the multivariate analysis the associations between education and respiratory health attenuated after adjusting for occupational exposure, smoking and outdoor air pollution. The crude odds ratio for the association between the lung function indicator FEV1 less than 80% of predicted value and educational level (<10 years vs. >10 years of school education) was 1.83 (95% CI: 1.22-2.74). This changed to 1.56 (95% CI: 1.03-2.37) after adjusting for occupational exposure, smoking and outdoor air pollution. CONCLUSION: We found an association between socio-economic status and respiratory health. This can partly be explained by living conditions indicated by occupational exposure, smoking behaviour and ambient air pollution. A relevant part of the social differences in respiratory health, however, remained unexplained.

Sunbed use induces the photoaging-associated mitochondrial common deletion.

Sunbeds have become very popular and, as a result, health effects of their use are of growing concern. In this regard, exposure to ultraviolet radiation from sunbeds may cause premature skin aging. Evidence for this assumption, however, has thus far been elusive. In this study, we assessed whether sunbed use induced the generation of a large-scale mitochondrial DNA mutation in human skin, the so-called Common Deletion, which is well known to be photoaging-associated. Fifty-nine individuals who voluntarily started to use sunbeds were observed over 3 months. Punch biopsies were taken from neck and buttock skin before the first and after their last sunbed visit. The Common Deletion content was determined by quantitative real-time PCR. Multiple linear regression analysis was performed for statistical analysis. Volunteers without previous sunbed use showed a 2.56-fold increase (95% CI: 0.97-6.78) of the Common Deletion in neck and a 3.58-fold increase (95% CI: 1.44-8.89) of the Common Deletion in buttock skin samples after sunbed use. Participants with previous sunbed use showed no increase. This study indicates that sunbed use may contribute to mtDNA deletion-associated skin photoaging.