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• W7148314 abstract "Engineered adipose tissue equivalents are increasingly acknowledged to serve for both fat grafting and basic research of obesity. As the current approaches do not allow for the formation of the desired coherent constructs, this thesis focused on the establishment of a fat-like model system capable of overcoming the present limitations. The 3-D model system was composed of the preadipocyte cell line 3T3-L1 and commercially available polyglycolic acid (PGA) fiber meshes as polymeric cell carrier. To assess appropriate tissue-inducing substances, different cell culture media and hormonal induction protocols (including the PPARgamma agonists indomethacin and troglitazone) were evaluated. The results of the study indicated that adipose conversion of 3T3-L1 cells was substantially improved in alpha-MEM as compared to DMEM. Furthermore, hormonal induction with corticosterone, IBMX, insulin, and indomethacin resulted in appropriate differentiation properties suitable for the TE of fat. In order to assemble fat-like constructs, distinct static and dynamic cultivation conditions were evaluated. In detail, PGA fiber meshes were dynamically seeded with 3T3-L1 preadipocytes; subsequently, cell-polymer constructs were hormonally induced and cultivation was performed either statically or dynamically in well-plates or in stirred bioreactors. Thereby, it could be shown that static cultivation in well plates is inferior to dynamic conditions with regard to the generation of adipose tissue. As the bioreactor approach did not feature any advantages over dynamic cultivation in well plates but entailed enhanced experimental requirements, it was considered to be less suitable to be routinely applied. The established cultivation conditions were employed for the generation of fat-like constructs, which were characterized extensively. After short-term culture over 9 days, the engineered constructs featured tissue-like coherence and exhibited typical characteristics of adipocytes. However, die-punching showed that they were composed of well-differentiated cells in outer areas and less differentiated ones in internal parts. Furthermore, as compared to native fat, the adipocytes had not yet adopted the typical signet-ring form of unilocular cells. Therefore, the constructs were differentiated for prolonged periods of time (up to 35 days). Analysis of the resulting tissues elucidated completion of reorganization to structures histologically comparable to those of native adipose tissue. The microscopic observations were supported by investigations on the cellular and the molecular level. Afterwards, the engineered constructs were examined under physiological conditions in vivo. Not only were differentiated constructs s.c. implanted into nude mice but also PGA meshes seeded with undifferentiated 3T3-L1 preadipocytes. Subsequent to their excision, it could be shown that 3T3-L1 cells, when transplanted in the form of differentiated TE constructs, yielded vascularized fat pads histologically comparable to native fat. In contrast, blank control meshes and preadipocyte-seeded scaffolds did not result in adipose tissue formation in vivo.Finally, Me.PEG-PLAs diblock copolymers, recently developed for controlled cell-biomaterial interactions, were investigated with regard to their usefulness for adipose tissue engineering. The biomaterials were processed into both 2-D polymer films and 3-D scaffolds, which were sterilized by UV irradiation prior to contact with the cells. In order to ensure unchanged polymer properties, an appropriate duration of exposure to UV light had to be assessed. For this purpose, films were subjected to UV irradiation for different periods of time and subsequently polymer properties were examined. The results indicated that UV treatment for 2 hours is an appropriate technique for sterilization of the Me.PEG-PLAs. However, UV irradiation for extended periods of time (5 and more hours) drastically altered the properties of the polymer films. Hence, adipogenesis on Me.PEG-PLAs was studied on cell carriers, which were previously sterilized by exposure to UV light for 2 hours. Me.PEG2-PLA20 polymers were demonstrated to allow for adipogenesis of 3T3-L1 cells. Nevertheless, the data from this study did not reveal an advantage over PGA or PLGA polymers with respect to adipose tissue development.In summary, this thesis for the first time demonstrated the development of a coherent fat-like tissue in vitro. Furthermore, it could be shown, that s.c. implantation of these constructs into nude mice allows for development of vascularized fat pads in vivo. Hence, TE was proven useful to establish a valuable tool for investigation of adipogenesis within a tissue-like environment in vitro and in vivo. Application of the model is suggested to elucidate tissue inherent interactions and, thus, to contribute to a better understanding of adipose tissue physiology and development of obesity." @default.
• W7148314 created "2016-06-24" @default.
• W7148314 creator A5065389019 @default.
• W7148314 date "2004-04-12" @default.
• W7148314 modified "2023-09-27" @default.
• W7148314 title "Adipose tissue engineering. Development of a 3-D model system of adipogenesis" @default.
• W7148314 hasPublicationYear "2004" @default.
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