Evidence for in vitro acinar-to-ductal transdifferentiation of human pancreas

Isabelle Houbracken, Evelien De Waele, Jessy Lardon, Zhidong Ling, Henry Heimberg, Luc Bouwens

Onderzoeksoutput: Meeting abstract (Book)

Samenvatting

Specific aims: To optimize and apply a system for genetic lineage tracing of human acinar cells in vitro and to show evidence for the plasticity of these cells.

Relevance to program: A better understanding of the plasticity of human exocrine cells might be important for finding a treatment for diabetes, since rodent acinar cells have been shown to be able to transdifferentiate to beta cells and to normalize alloxan-induced hyperglycemic mice.

General outline:
The plasticity of rodent pancreatic acinar cells was demonstrated in different in vitro and in vivo models, eg. our group showed that rat acinar cells transdifferentiate in vitro to beta cells via an intermediate duct-like phenotype. Controversy exists about the plasticity of human acinar cells since none of the observations have been confirmed via tracing. We created adenoviral vectors to deliver Amylase-nlsCre and CMV-Lox-STOP-Lox-EGFP to the human cell preparations to trace the acinar transdifferentiation genetically. Upon transfection of both constructs into AR42J, a rat exocrine cell line, we noticed a lot of bright EGFP+ cells; whereas transfection of the lox-construct only, resulted in very few, weak EGFP+ cells, indicating correct functioning, cre-mediated recombination. We have optimized the adenoviral transduction of human exocrine cells. When transduced on day 1 with Ad-EGFP, the labeling efficiency was 24,7 ± 2,4% (n=7) and 40,1 ± 4,9% (n=6) with multiplicity of infection (MOI) 5 and 10 respectively. When we applied both Ad-Amylase-nlsCre and Ad-CMV-Lox-STOP-Lox-EGFP on day 1, bright EGFP+ cells appeared in the culture, consistent with the results in AR42J. Quantification on day 7 showed 3,4 ± 0,4% EGFP+ cells (n=6). 93,6 ± 0,8% of these EGFP+ cells expressed the ductal marker cytokeratin19 on day 7, whereas only 0,83 ± 0,04% of them expressed the ductal marker on day 2. At this timepoint 99,5 ± 0,4% of the EGFP+ cells were chymotrypsin+, showing their acinar phenotype in the beginning of the culture. These results show evidence for the transdifferentiation of acinar cells to ductal cells.
Originele taal-2English
TitelPoster presentation BetaCellTherapy workshop Brussels 2010
StatusPublished - 21 jan 2010
EvenementUnknown -
Duur: 21 jan 2010 → …

Conference

ConferenceUnknown
Periode21/01/10 → …

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