Proteomics

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Omics analyses of 3T3-J2 feeder cells to identify culture conditions that support the long-term culture of human pancreatic progenitor cells


ABSTRACT: Multipotent pancreatic progenitor cells have been successfully cultured, expanded and differentiated into cells of the adult human pancreas. The long-term culture of these progenitors (termed cPP) relies on support from mitotically inactivated 3T3-J2 mouse embryonic fibroblasts. The 3T3-J2-derived signals that support cPP expansion and maintain the fine balance between self-renewal and lineage commitment in this co-culture system remain unknown. To unmask these signals, we exploited a striking disparity in morphology and the capacity to self-renew of cPP cultured on 3T3-J2 cells from different sources. We deployed a comparative approach to identify (1) genes that define expandable cPP through transcriptome analysis and (2) key factors secreted by 3T3-J2 using selective enrichment of newly synthesised proteins and mass spectrometry analysis. Using cPP from three genetic backgrounds cultured on either supportive or non-supportive 3T3-J2, these analyses revealed 100 transcription factor genes that are significantly down-regulated in cPP cultured on non-supportive 3T3-J2. Analysis of differentially expressed 3T3-J2 proteins identified 53 whose expression is higher in supportive 3T3-J2. These results provide significant insight into the requirements not only for optimal cPP culture and expansion but also for progenitor cell types in other organ systems that rely on 3T3-J2 co-culture, most notably keratinocytes for clinical applications.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Norris Ray Dunn 

PROVIDER: PXD041758 | JPOST Repository | Wed Apr 24 00:00:00 BST 2024

REPOSITORIES: jPOST

Dataset's files

Source:
Action DRS
20190706_RS_Jamie_1.msf Msf
20190706_RS_Jamie_1.pdResult Other
20190706_RS_Jamie_1.xlsx Xlsx
20190706_RS_Jamie_10_1.raw Raw
20190706_RS_Jamie_10_2.raw Raw
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