Longitudinal scRNA-seq analysis in mouse and human informs optimization of rapid mouse astrocyte differentiation protocols.

Macroglia (astrocytes and oligodendrocytes) are required for normal development and function of the central nervous system, yet many questions remain about their emergence during the development of the brain and spinal cord. Here we used single-cell/single-nucleus RNA sequencing (scRNA-seq/snRNA-seq) to analyze over 298,000 cells and nuclei during macroglia differentiation from mouse embryonic and human-induced pluripotent stem cells. We computationally identify candidate genes involved in the fate specification of glia in both species and report heterogeneous expression of astrocyte surface markers across differentiating cells. We then used our transcriptomic data to optimize a previous mouse astrocyte differentiation protocol, decreasing the overall protocol length and complexity. Finally, we used multi-omic, dual single-nuclei (sn)RNA-seq/snATAC-seq analysis to uncover potential genomic regulatory sites mediating glial differentiation. These datasets will enable future optimization of glial differentiation protocols and provide insight into human glial differentiation.

Biomechanical evaluation of parasagittal occipital plating: screw load sharing analysis.

STUDY DESIGN: Biomechanical evaluation of occipitocervical instrumentation techniques. OBJECTIVE: Compare methods of occipital instrumentation by quantifying load sharing of occipital screws and measuring motion across instrumented occipitocervical spines. SUMMARY OF BACKGROUND DATA: Newer occipitocervical plate/screw systems that attach to longitudinal rods have been developed to improve fixation. These devices place screws in the center of occipital bone or off-midline. Midline plates offer screw purchase in thicker bone. Off-midline systems may increase the effective moment arm for torsional and lateral bending control. Measurement of screw loads within occipital plates is useful for determining optimal plate configuration. METHODS: Ten cadaveric specimens (occiput-C4) were tested in flexion/extension (FE), lateral bending (LAT), and axial rotation (ROT) over +/-3 Nm pure moment. After intact testing, 4 occipitocervical fixation constructs were tested using washer load cells to assess loading across screws used to fix the plates to the occiput. Parasagittal occipital plates were positioned either convex or concave side facing medially. Each plate was first fixed using 3 screws (rostral, middle, caudal), then with the caudal screw eliminated (simulated failure). Range of motion (ROM) and peak screw loads are reported. RESULTS: ROM decreased from intact to any of the 4 fusion plate configurations in FE, LAT, and ROT (P << 0.05), but not between plate configurations. Screw load significantly decreased from medially convex to medially concave configurations in LAT, but no significant changes were observed in FE or ROT. With caudal screws removed, middle screws peak loads significantly increased in FE and LAT (P < 0.05), but not ROT. CONCLUSION: Occipital screw placement off-midline improves screw loads under lateral bending forces on occipitocervical constructs, though loads for FE and ROT are unchanged. As screws pullout, the loads may be redistributed, resulting in increased screw pullout forces above. Despite the improvement in screw loads for laterally based plates during lateral bending, overall ROM across the occipitocervical junction is unchanged.