Project description:Adult neural progenitor cells (aNPCs) are a potential source for cell based therapy for neurodegenerative diseases and traumatic brain injuries. We show that the ultrasonic aspirate samples that are typically considered as a waste after surgery are a great source for aHNPCs.
Project description:Single-nucleus RNA sequencing (snRNA-seq) was used to profile the transcriptome of 16,015 nuclei in human adult testis. This dataset includes five samples from two different individuals. This dataset is part of a larger evolutionary study of adult testis at the single-nucleus level (97,521 single-nuclei in total) across mammals including 10 representatives of the three main mammalian lineages: human, chimpanzee, bonobo, gorilla, gibbon, rhesus macaque, marmoset, mouse (placental mammals); grey short-tailed opossum (marsupials); and platypus (egg-laying monotremes). Corresponding data were generated for a bird (red junglefowl, the progenitor of domestic chicken), to be used as an evolutionary outgroup.
Project description:Robust and reliable and in vitro models are essential in the discovery of new treatment options for high-grade glioma (HGG), however, establishing successful patient-derived cell cultures has posed significant challenges. We established glioma stem-like cell (GSC) cultures from 114 consecutive HGG specimens, obtained via traditional surgical resection and/or ultrasonic aspiration, using fully-dissociated single cells (single cell-derived, SCD) and partially dissociated tissue fragments (3D-derived, 3DD). Copy number profiling assessed genetic similarities, while single-cell RNA sequencing evaluated transcriptomic heterogeneity. Proof-of-concept personalized drug screening was performed using a panel of approved anti-cancer agents. Higher success rates in establishing GSC cultures were obtained from ultrasonic aspirates (SCD and 3DD) and 3DD surgical samples compared to SCD cultures from surgical samples. Combining these approaches in parallel yielded a 96% success rate. In rare cases, copy number variations in original tumor tissue were lost in culture, leading to discernible changes in cell morphology. Single-cell sequencing revealed greater heterogeneity of transcriptomic clusters in ultrasonic aspiration-derived cultures compared to resection-derived cultures. Our study's protocol supported the screening of 20 anti-cancer agents within a clinically-relevant timeframe in 16 of 18 consecutive HGG samples. Our optimized protocol therefore provides a reliable tool for generating HGG cell cultures which capture the tumors’ molecular characteristics and supports precision medicine applications.
Project description:Robust and reliable and in vitro models are essential in the discovery of new treatment options for high-grade glioma (HGG), however, establishing successful patient-derived cell cultures has posed significant challenges. We established glioma stem-like cell (GSC) cultures from 114 consecutive HGG specimens, obtained via traditional surgical resection and/or ultrasonic aspiration, using fully-dissociated single cells (single cell-derived, SCD) and partially dissociated tissue fragments (3D-derived, 3DD). Copy number profiling assessed genetic similarities, while single-cell RNA sequencing evaluated transcriptomic heterogeneity. Proof-of-concept personalized drug screening was performed using a panel of approved anti-cancer agents. Higher success rates in establishing GSC cultures were obtained from ultrasonic aspirates (SCD and 3DD) and 3DD surgical samples compared to SCD cultures from surgical samples. Combining these approaches in parallel yielded a 96% success rate. In rare cases, copy number variations in original tumor tissue were lost in culture, leading to discernible changes in cell morphology. Single-cell sequencing revealed greater heterogeneity of transcriptomic clusters in ultrasonic aspiration-derived cultures compared to resection-derived cultures. Our study's protocol supported the screening of 20 anti-cancer agents within a clinically-relevant timeframe in 16 of 18 consecutive HGG samples. Our optimized protocol therefore provides a reliable tool for generating HGG cell cultures which capture the tumors’ molecular characteristics and supports precision medicine applications.
Project description:Transcriptome analysis of neural progenitor/stem cells is limited by the lack of a reliable method for cell isolation. We have designed a genetic dual reporter strategy that can allow prospective isolation of cortical neural progenitor cells and their neuronal progeny form the same animals. These cells should be a good cell source for comparative global analysis.
Project description:Transcriptome analysis of neural progenitor/stem cells is limited by the lack of a reliable method for cell isolation. We have designed a genetic dual reporter strategy that can allow prospective isolation of cortical neural progenitor cells and their neuronal progeny form the same animals. These cells should be a good cell source for comparative global analysis. Cortical neural progenitor cells and their neuronal progeny were purified using a dual reporter strategy. The purified cells were used for comparative expression profiling.
Project description:RNA-seq samples from 3 species across a differentiation from induced pluripotent stem cells to neural progenitor cells were generated to study gene expression evolution. Briefly, previously generated urinary stem cell derived iPSCs of 3 human (Homo sapiens) individuals (3 clones), 1 gorilla (Gorilla gorilla) individual and fibroblast derived cynomolgus macaque (Macaca fascicularis) iPSCs of 2 individuals (4 clones) (Geuder et al. 2021) were differentiated to neural progenitor cells via dual-SMAD inhibition as three-dimensional aggregation culture (Chambers et al. 2009; Ohnuki et al. 2014). Bulk RNA-seq libraries of iPSCs and NPCs were generated using prime-seq protocol (Janjic et al. 2022).