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Single-cell multimodal approaches and machine learning are transforming analysis of the human hematopoietic progenitor compartment. Cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) enables coupled surface protein and transcriptome profiling, revealing genomic programs underlying progenitor states.

This webcast will present groundbreaking work establishing a high-resolution human bone marrow single-cell atlas using paired CITE-seq and flow cytometry, offering an integrated genomic, bioinformatics and flow cytometric resource for diverse hematopoietic progenitor cell states in human health and disease.

The new study systematically performed CITE-seq on primary human bone marrow cells using titrations with 266 antibody derived tags (ADTs) to develop a custom, 132-plex optimized cocktail. Curiox Laminar Wash technology streamlined ADT and hashtag antibody staining/washing with great reproducibility and minimal background.

Applying the optimized cocktail on expanded cohorts revealed over 80 distinct cell types, including stem, progenitor, immune and stromal cells defined by distinctive surface markers and transcriptomes. Laminar Wash Direct Reading Grid empowered high throughput full-spectrum cytometry profiling the same 132 surface markers in parallel with CITE-seq, and collaboratively nominated novel and known markers to segregate transitional cell states.

Integrative analysis of CITE-seq and flow cytometry identified dozens of markers consistently detected across donors spanning race and sex. Aligning annotations from this atlas nominated normal marrow equivalents for acute myeloid leukemia stem cell populations that differ in clinical response.

• The process and benefits of using CITE-seq and high-dimensional flow cytometry techniques on bone marrow samples
• How to best process samples to optimize ADT and hashtag antibody staining/washing
• How to achieve purer cells in a sorting gate using differential markers