Changes in gene expression and morphology of mouse embryonic stem cells on differentiation into insulin-producing cells in vitro and in vivo

Autor: Flavio Francini, Sally Picton, Sigurd Lenzen, Ortwin Naujok, Clifford J. Bailey, Anne Jörns
Rok vydání: 2009
Předmět:
Male
medicine.medical_specialty
Amyloid
CIENCIAS MÉDICAS Y DE LA SALUD
Endocrinology
Diabetes and Metabolism
Cellular differentiation
Cell Culture Techniques
Nerve Tissue Proteins
Medicina Clínica
Biology
Diabetes Mellitus
Experimental
Cell therapy
Nestin
Mice
Endocrinology
Intermediate Filament Proteins
In vivo
Internal medicine
Endocrinología y Metabolismo
Insulin-Secreting Cells
Glucokinase
Internal Medicine
medicine
Animals
Insulin
INSULIN CELL THERAPY
INSULIN-PRODUCING CELLS
Embryonic Stem Cells
Glucose Transporter Type 2
Homeodomain Proteins
MOUSE EMBRYONIC STEM CELLS
Gene Expression Profiling
Cell Differentiation
Embryonic stem cell
Cell biology
Islet Amyloid Polypeptide
DIFFERENTIATION
Diabetes Mellitus
Type 1
Gene Expression Regulation
Cell culture
Trans-Activators
PDX1
Stem cell
Biomarkers
Stem Cell Transplantation
Zdroj: Diabetes/metabolism research and reviews. 25(5)
ISSN: 1520-7560
Popis: Background: Embryonic stem (ES) cells have the potential to produce unlimited numbers of surrogate insulin-producing cells for cell replacement therapy of type 1 diabetes mellitus. The impact of the in vivo environment on mouse ES cell differentiation towards insulin-producing cells was analysed morphologically after implantation. Methods: ES cells differentiated in vitro into insulin-producing cells according to the Lumelsky protocol or a new four-stage differentiation protocol were analysed morphologically before and after implantation for gene expression by in situ reverse transcription polymerase chain reaction and protein expression by immunohistochemistry and ultrastructural analysis. Results: In comparison with nestin positive ES cells developed according to the reference protocol, the number of ES cells differentiated with the four-stage protocol increased under in vivo conditions upon morphological analysis. The cells exhibited, in comparison to the in vitro situation, increased gene and protein expression of Pdx1, insulin, islet amyloid polypeptide (IAPP), the GLUT2 glucose transporter and glucokinase, which are functional markers for glucose-induced insulin secretion of pancreatic beta cells. Renal sub-capsular implantation of ES cells with a higher degree of differentiation achieved by in vitro differentiation with a four-stage protocol enabled further significant maturation for the beta-cell-specific markers, insulin and the costored IAPP as well as the glucose recognition structures. In contrast, further in vivo differentiation was not achieved with cells differentiated in vitro by the reference protocol. Conclusions: A sufficient degree of in vitro differentiation is an essential prerequisite for further substantial maturation in a beta-cell-specific way in vivo, supported by cell-cell contacts and vascularisation. Fil: Naujok, Ortwin. Hannover Medical School; Alemania Fil: Francini, Flavio. Hannover Medical School; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina Fil: Picton, Sally. Aston University; Reino Unido Fil: Bailey, Clifford J.. Aston University; Reino Unido Fil: Lenzen, Sigurd. Hannover Medical School; Alemania Fil: Jörns, Anne. Hannover Medical School; Alemania
Databáze: OpenAIRE
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