Standardized generation of human iPSC-derived hematopoietic organoids and macrophages utilizing a benchtop bioreactor platform under fully defined conditions.

BACKGROUND: There is a significant demand for intermediate-scale bioreactors in academic and industrial institutions to produce cells for various applications in drug screening and/or cell therapy. However, the application of these bioreactors in cultivating hiPSC-derived immune cells and other blood cells is noticeably lacking. To address this gap, we have developed a xeno-free and chemically defined intermediate-scale bioreactor platform, which allows for the generation of standardized human iPSC-derived hematopoietic organoids and subsequent continuous production of macrophages (iPSC-Mac). METHODS: We describe a novel method for intermediate-scale immune cell manufacturing, specifically the continuous production of functionally and phenotypically relevant macrophages that are harvested on weekly basis for multiple weeks. RESULTS: The continuous production of standardized human iPSC-derived macrophages (iPSC-Mac) from 3D hematopoietic organoids also termed hemanoids, is demonstrated. The hemanoids exhibit successive stage-specific embryonic development, recapitulating embryonic hematopoiesis. iPSC-Mac were efficiently and continuously produced from three different iPSC lines and exhibited a consistent and reproducible phenotype, as well as classical functionality and the ability to adapt towards pro- and anti-inflammatory activation stages. Single-cell transcriptomic analysis revealed high macrophage purity. Additionally, we show the ability to use the produced iPSC-Mac as a model for testing immunomodulatory drugs, exemplified by dexamethasone. CONCLUSIONS: The novel method demonstrates an easy-to-use intermediate-scale bioreactor platform that produces prime macrophages from human iPSCs. These macrophages are functionally active and require no downstream maturation steps, rendering them highly desirable for both therapeutic and non-therapeutic applications.

The prevalence of anabolic androgenic steroid use amongst athletes in Riyadh (Saudi Arabia).

OBJECTIVE: The aim of this study was to determine the prevalence of anabolic androgenic steroid (AAS) use among athletes and examine the extent of their knowledge on the effects of AAS in Riyadh, Saudi Arabia. METHODS: This cross-sectional study was conducted at gyms in Riyadh, Saudi Arabia, during 2015. In total 600 athletes from three gyms participated in the study. The study included Saudi and non-Saudi athletes chosen by the simple random sampling method. A self-reported questionnaire was used for data collection. The questionnaire was designed to study the prevalence and assess the knowledge of athletes regarding AAS use. Frequency and percentage distributions were used to describe the data. Comparison between the subgroups was made with a chi-square test. RESULTS: The percentage of AAS users was 30.5%. The age of AAS users ranged from 15 to 49 years with the majority (52.5%) belonging to age group of 25-29 years. Approximately 20% of the users admitted using AAS due to body dysmorphia as their best motivational factor; in addition, they also believed that there are no side effects of the use. Among the nonusers, 40% had appropriate knowledge, while all the AAS-users had inadequate knowledge about the adverse effects of AAS. Moreover, 77% of the users would recommend AAS to their friends but none from the nonusers. A significant difference in age distribution (df = 5, p<0.001) and knowledge (df = 4, p< 0.001) between users and nonusers was observed. CONCLUSION: Most athletes were ignorant of the harmful side effects of the drug but still continued to use and promote it to other athletes. These athletes should intensify their knowledge and awareness regarding the use of AAS and its effects on the body.