Adult human pancreas-derived cells expressing stage-specific embryonic antigen 4 differentiate into Sox9-expressing and Ngn3-expressing pancreatic ducts in vivo

Abstract

BackgroundTissue-specific stem/progenitor cells are found in various adult tissues and may have the capacity for lineage-specific differentiation, facilitating applications in autologous transplantation. Stage-specific embryonic antigen 4 (SSEA-4), an early embryonic glycolipid antigen, is expressed in cells derived from adult human pancreas exocrine tissue. Here, we examined the characteristics and lineage-specific differentiation capacity of SSEA-4+ cells.MethodsHuman adult partial pancreas tissues were obtained from different donors and cultured in vitro. SSEA-4+ and CA19-9+ cells were isolated from adult human pancreas exocrine cells using magnetic-activated cell sorting, and gene expression was validated by quantitative polymerase chain reaction. To confirm in-vivo differentiation, SSEA-4+ and CA19-9+ cells were transplanted into the dorsal subcutaneous region of mice. Finally, morphological features of differentiated areas were confirmed by immunostaining and morphometric analysis.ResultsSSEA-4-expressing cells were detected in isolated pancreas exocrine cells from adult humans. These SSEA-4+ cells exhibited coexpression of CA19-9, a marker of pancreatic duct cells, but not amylase expression, as shown by immunostaining and flow cytometry. SSEA-4+ cells exhibited higher relative expression of Oct4, Nanog, Klf4, Sox2, and c-Myc mRNAs than CA19-9+ cells. Pancreatic intralobular ducts (PIDs) were generated from SSEA-4+ or CA19-9+ cells in vivo at 5 weeks after transplantation. However, newly formed PIDs from CA19-9+ cells were less abundant and showed an incomplete PID morphology. In contrast, newly formed PIDs from SSEA-4+ cells were abundant in the transplanted area and showed a crowded morphology, typical of PIDs. Sox9 and Ngn3, key transcription factors associated with pancreatic development and regeneration, were expressed in PIDs from SSEA-4+ cells.ConclusionsSSEA-4-expressing cells in the adult human pancreas may have the potential for regeneration of the pancreas and may be used as a source of stem/progenitor cells for pancreatic cell lineage-specific differentiation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0422-0) contains supplementary material, which is available to authorized users.

Role of c-MET Inhibitors in Overcoming Drug Resistance in Spheroid Models of Primary Human Pancreatic Cancer and Stellate Cells

“…The resulting primary cultures were transduced with lentiviral vectors encoding firefly luciferase and CFP, as described previously [56,57]. PDAC5 cells were sorted for stage-specific embryonic antigen-4 (SSEA4) positive cells by flow cytometry to generate the PDAC5 (SSEA4) subclone [58].…”

The Role of Krüppel-like Factors in Pancreatic Physiology and Pathophysiology

“…Purification and expansion of native pancreatic stem cells have been attempted and failed because they comprise such a small proportion of the pancreatic cell population [60]. Deriving and reprogramming adult human pancreas cells expressing embryonic antigen 4, showed that tissue-specific/progenitor cells have an excellent capacity for lineagespecific differentiation, which could facilitate autologous transplantations [63]. These cells exhibited higher expression of KLF4, SOX2, OCT4, and homeobox transcription factor Nanog (NANOG) mRNA levels, suggesting greater potential as a source of inducible pancreatic lineage-specific cells compared to other previously used cell types such as fibroblasts [63].…”

“…Deriving and reprogramming adult human pancreas cells expressing embryonic antigen 4, showed that tissue-specific/progenitor cells have an excellent capacity for lineagespecific differentiation, which could facilitate autologous transplantations [63]. These cells exhibited higher expression of KLF4, SOX2, OCT4, and homeobox transcription factor Nanog (NANOG) mRNA levels, suggesting greater potential as a source of inducible pancreatic lineage-specific cells compared to other previously used cell types such as fibroblasts [63]. Additionally, positive correlations have been found between the number of induced pancreatic islet-specific progenitors transplanted and the long-term post-transplant metabolic success of glucose tolerance [60].…”