Ultrastructural and molecular analyzes of insulin-producing cells induced from human hepatoma cells.
OBJECTIVE
Diabetes type I is an autoimmune disease characterized by the destruction of pancreatic insulin-producing (beta-) cells and resulting in external insulin dependence for life. Islet transplantation represents a potential treatment for diabetes but there is currently a shortage of suitable organs donors. To augment the supply of donors, different strategies are required to provide a potential source of beta-cells. These sources include embryonic and adult stem cells as well as differentiated cell types. The main goal of this study was to induce the transdifferentiation (or conversion of one type cell to another) of human hepatoma cells (HepG2 cells) to insulin-expressing cells based on the exposure of HepG2 cells to an extract of rat insulinoma cells (RIN).


METHODS
HepG2 cells were first transiently permeabilized with Streptolysin O and then exposed to a cell extract obtained from RIN cells. Following transient exposure to the RIN extract, the HepG2 cells were cultured for 3 weeks.


RESULTS
Acquisition of the insulin-producing cell phenotype was determined on the basis of (i) morphologic and (ii) ultrastructural observations, (iii) immunologic detection and (iv) reverse transcription (RT)-polymerase chain reaction (PCR) analysis.


CONCLUSIONS
This study supports the use of cell extract as a feasible method for achieve transdifferentiation of hepatic cells to insulin-producing cells.
Perán
Macarena
M
Department of Health Sciences, University of Jaén, Jaén, Spain.
Sánchez-Ferrero
Aitor
A
Tosh
David
D
Marchal
Juan Antonio
JA
Lopez
Elena
E
Alvarez
Pablo
P
Boulaiz
Houria
H
Rodríguez-Serrano
Fernando
F
Aranega
Antonia
A
D000818:Animals; D001426:Bacterial Proteins; D002457:Cell Extracts / Q000494:pharmacology; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D056945:Hep G2 Cells / Q000166:cytology*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000096:biosynthesis*; D050417:Insulin-Secreting Cells / Q000378:metabolism* / Q000648:ultrastructure*; D007340:Insulinoma; D046529:Microscopy, Electron, Transmission; D010539:Permeability; D010641:Phenotype; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013301:Streptolysins
D001426:Bacterial Proteins; D002457:Cell Extracts; D007328:Insulin; D013301:Streptolysins; C025964:streptolysin O