Artikel
Generation of insulin producing cells from non human primate monocytes derived from an optimized protocol for the generation of human neo-islet cells
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Veröffentlicht: | 20. Mai 2011 |
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Introduction: According to a previously published protocol for the generation of Insulin-producing cells from human monocytes we now analyzed wether the addition of 4 different growth factors could optimize Insulin production.
Secondly we tried to generate these cells from non-human primate monocytes either blood or spleen derived.As a preliminary experiment for a large animal model of induced diabetes and potential autologous treatment we generated Insulin producing cells from a complete spleen after splenectomy of a live baboon (Papio anubis).
Materials and methods: Human monocytes were treated according to a published Protocol (Ruhnke et al., Gastroenterology 2005) to dedifferenciate into PCMO (Programmable cells of monocytic origin).
In addition to the previously published differentiation protocoll PCMO's then received treatment with either activin a, betacellulin, exendin 3 or 4. Cells were analyzed according to the expression of Insulin, PDX-1, C-peptide, Glut-2 and insulin content.
After identifying the optimal protocoll, we isolated monocytes from baboon blood and repeated the protocol. Furthermore a fully grown female baboon was anethetized and unterwent median laparotomy with subsequent splenectomy. Spleen monocytes were differentiated and analyzed in the same manner and calculated in number and volume for potential further in vivo autologous application.
Results: Insulin content of human neo-islets was highest when cells were treated with activin a addition. Insulin content was 13,000 µU/1 Million cells.
This protocoll was used fro the differentiation of primate monocytes. Surprisingly we could generate Insulin producing neo-islets cells form primate monocytes despite using only human growth factors and serum.
Expresson of Insulin, PDX-1, C-peptide and Glut-2 was comparable to that of human neo-islets.
Total insilin content of activin a treated baboon monocytes was comparable to that of human neo-Islets with appr. 16,000 µU/1Million cells.
Conclusion: We could show that Insulin producing cells could be generated from baboon monocytes with human growth factors. The amount generated from one spleen could be enough to cure a 40kg baboon from an experimentally induced diabetes in an autologous cell transplant setting.