Figure 4.

Droplet-Based Single-Cell PCR Analysis of Peripheral Blood and Bone Marrow Samples from Patients

Droplet-based single-cell PCR (dsPCR) was performed using samples from two patients. (A) Immunological characterization of peripheral blood and bone marrow samples from patients via FACS analysis. Percentages of CD3⁺ T cells, CD56⁺ NK cells, and CD19⁺ B cells in lymphocytes are shown in the bar chart. Bone marrow samples were also analyzed for CD34 expression. (B) Ratios of vector-positive cells in the whole PBMC and sorted fractions. PBMC samples from patients were sorted via FACS into CD3⁺ T cell, CD56⁺ NK cell, and CD19⁺ B cell fractions, and each cell fraction and all PBMCs were encapsulated into droplets. The ratio of vector-positive cells was determined by single cell-based digital droplet PCR (sc-ddPCR). Data were pooled from three independent experiments for each patient. (C) Bone marrow samples after lysis of red blood cells were also sorted and analyzed for the presence of the vector ψ sequence (whole bone marrow cells, CD34⁺ cells). For each sample, sc-ddPCR was conducted in triplicate.