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Comparative Study
. 2011 Nov;54(11):2832-44.
doi: 10.1007/s00125-011-2283-5. Epub 2011 Sep 1.

Transcriptomes of the major human pancreatic cell types

C Dorrell  1 , J Schug, C F Lin, P S Canaday, A J Fox, O Smirnova, R Bonnah, P R Streeter, C J Stoeckert Jr, K H Kaestner, M Grompe
Affiliations
Comparative Study

Transcriptomes of the major human pancreatic cell types

C Dorrell et al. Diabetologia. 2011 Nov.

Erratum in

  • Diabetologia. 2013 May;56(5):1192

Abstract

Aims/hypothesis: We sought to determine the mRNA transcriptome of all major human pancreatic endocrine and exocrine cell subtypes, including human alpha, beta, duct and acinar cells. In addition, we identified the cell type-specific distribution of transcription factors, signalling ligands and their receptors.

Methods: Islet samples from healthy human donors were enzymatically dispersed to single cells and labelled with cell type-specific surface-reactive antibodies. Live endocrine and exocrine cell subpopulations were isolated by FACS and gene expression analyses were performed using microarray analysis and quantitative RT-PCR. Computational tools were used to evaluate receptor-ligand representation in these populations.

Results: Analysis of the transcriptomes of alpha, beta, large duct, small duct and acinar cells revealed previously unrecognised gene expression patterns in these cell types, including transcriptional regulators HOPX and HDAC9 in the human beta cell population. The abundance of some regulatory proteins was different from that reported in mouse tissue. For example, v-maf musculoaponeurotic fibrosarcoma oncogene homologue B (avian) (MAFB) was detected at equal levels in adult human alpha and beta cells, but is absent from adult mouse beta cells. Analysis of ligand-receptor interactions suggested that EPH receptor-ephrin communication between exocrine and endocrine cells contributes to pancreatic function.

Conclusions/interpretation: This is the first comprehensive analysis of the transcriptomes of human exocrine and endocrine pancreatic cell types-including beta cells-and provides a useful resource for diabetes research. In addition, paracrine signalling pathways within the pancreas are shown. These results will help guide efforts to specify human beta cell fate by embryonic stem cell or induced pluripotent stem cell differentiation or genetic reprogramming.

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Figures

Fig. 1
Fig. 1
The isolation of subsets of live human pancreatic endocrine and exocrine cells. a The FACS-detected fluorescence of pancreatic islet cells co-labelled with endocrine cell markers HPi2 and HPa2, with back-scattering visualisation of (b) beta and (c) alpha cells to reveal their size/granularity (FSC/side scatter [SSC]) characteristics. d As above (a), but co-labelled with exocrine markers HPx1, HPd1 and HPd3, and with back-scattering visualisation of the populations defined as acinar (e), large duct (f) and small duct (g). h Dual immunofluorescent labelling of adult human pancreatic cryosections simultaneously labelled with the same (a, b) groups of endocrine or (i, j) exocrine cell type-specific antibodies, respectively. k The comprehensive duct labelling of HPd3 is demonstrated by co-labelling with KRT19. l The relative levels of markers of beta (insulin), alpha (glucagon), delta (somatostatin), acinar (PRSS1) or duct (KRT19) cell identity. Quantitative RT-PCR results obtained from FACS-isolated populations were calculated as ΔCt values relative to the mean of Lamin A/C and 18S rRNA. The total signal detected for each marker gene is indicated as a percentage
Fig. 2
Fig. 2
Hierarchical clustering of microarray-assessed gene expression in human pancreatic endocrine and exocrine cell subpopulations. We used PaGE algorithm to identify 5,038 differentially expressed genes (in any pair-wise comparison; 95% CI), and clustered them hierarchically using TIGR Multiexperiment Viewer
Fig. 3
Fig. 3
Gene expression distribution of known and novel transcriptional regulators in pancreatic cell subpopulations. a, b, f Quantitative RT-PCR results obtained from RNA isolated from FACS-isolated human pancreatic cell populations were calculated as ΔCt values relative to the mean of housekeeping genes Lamin A/C and 18S rRNA. The total signal detected for each gene in each pancreatic cell subpopulation is indicated in per cent. c–e Protein levels of HDAC9, HOPX and β-actin were determined by SDS-PAGE of lysates obtained from sorted populations and by western blotting
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