A new method for generating insulin-secreting cells from human pancreatic epithelial cells after islet isolation transformed by NeuroD1.
The generation of insulin-secreting cells from nonendocrine pancreatic epithelial cells (NEPEC) has been demonstrated for potential clinical use in the treatment of diabetes. However, previous methods either had limited efficacy or required viral vectors, which hinder clinical application. In this study, we aimed to establish an efficient method of insulin-secreting cell generation from NEPEC without viral vectors. We used nonislet fractions from both research-grade human pancreata from brain-dead donors and clinical pancreata after total pancreatectomy with autologous islet transplantation to treat chronic pancreatitis. It is of note that a few islets could be mingled in the nonislet fractions, but their influence could be limited. The NeuroD1 gene was induced into NEPEC using an effective triple lipofection method without viral vectors to generate insulin-secreting cells. The differentiation was promoted by adding a growth factor cocktail into the culture medium. Using the research-grade human pancreata, the effective method showed high efficacy in the differentiation of NEPEC into insulin-positive cells that secreted insulin in response to a glucose challenge and improved diabetes after being transplanted into diabetic athymic mice. Using the clinical pancreata, similar efficacy was obtained, even though those pancreata suffered chronic pancreatitis. In conclusion, our effective differentiation protocol with triple lipofection method enabled us to achieve very efficient insulin-secreting cell generation from human NEPEC without viral vectors. This method offers the potential for supplemental insulin-secreting cell transplantation for both allogeneic and autologous islet transplantation.
1 Baylor University Medical Center , Dallas, TX 75226.
Masayuki
M
Shimoda
Shuyuan
S
Chen
Hirofumi
H
Noguchi
Morihito
M
Takita
Koji
K
Sugimoto
Takeshi
T
Itoh
Daisuke
D
Chujo
Shuichi
S
Iwahashi
Bashoo
B
Naziruddin
Marlon F
MF
Levy
Shinichi
S
Matsumoto
Paul A
PA
Grayburn
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D002096:C-Peptide / Q000097:blood; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D003921:Diabetes Mellitus, Experimental / Q000628:therapy; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D005260:Female; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology*; D053539:Keratin-19 / Q000235:genetics; D008297:Male; D051379:Mice; D010179:Pancreas / Q000166:cytology; D011401:Promoter Regions, Genetic
D051792:Basic Helix-Loop-Helix Proteins; D002096:C-Peptide; D007328:Insulin; D053539:Keratin-19; C093970:NEUROD1 protein, human