Fig. 5 |. Human BM Chip recapitulates hematopoietic abnormalities observed in Shwachman-Diamond Syndrome patients.

a, Photographs of BM Chips seeded with CD34+ cells from normal donors versus SDS patients at 2 weeks of culture (n=4 chips per condition; representative of 2 independent experiments). Scale bar, 5mm. Neutrophil (b, left), erythroid (b, right), and CD34+ (c) cell numbers were quantified by flow cytometry. d, Percentages of neutrophils with a mature CD16^hi surface phenotype in control versus SDS BM Chips were quantified by flow cytometry. e, Graph showing the number of erythroid cells at different maturation states (left) and representative flow plots (right) depicting the percentages of the erythroid subpopulations (E1: immature, E3: mature), as quantified by flow cytometry. f, Representative flow plots depicting the abnormal CD13 vs CD16 neutrophil maturation pattern observed in SDS BM Chips (top) and SDS patient bone marrow aspirates (bottom) as compared to normal controls (SDS = blue, normal = gray). After gating on CD16^hi neutrophils in BM Chips and patient marrow samples, CD13 expression for SDS vs control samples was assessed as shown by the histograms (middle, each histogram represents a different patient) and quantified by comparing median fluorescence intensities (right, square and triangle data points belong to the 2 SDS patients tested in BM Chip cultures). (b-f, BM Chips: n=8, error bars represent s.d., data pooled from 2 independent experiments, each using cells from a different normal and SDS patient with 4 chips per experiment; Patient BM aspirates: n=4 normal and n=8 SDS patients; numbers indicate P values for normal versus SDS samples using two-tailed Student’s t-test except CD13 MFI levels for SDS patients fell into 2 distinct groups and were compared with that of control patients by Mann-Whitney U test; ***P < 0.001).