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Design and Fabrication of Micro-Fluidic Chip for Formation, Culture and Analysis of Cancer Tumor Cell Spheroids

Khoshravesh, Hirad | 2020

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 52685 (08)
  4. University: Sharif University of Technology
  5. Department: Mechanical Engineering
  6. Advisor(s): Saeedi, Mohammad Saeed; Kazemzadeh Hannani, Siamak
  7. Abstract:
  8. Simulating the in-vivo environment of cancer tumors outside the body, can provide an appropriate condition for studying different key processes of the tumor micro-environment, such as aggregation, formation, growth, angiogenesis and metastasis of the cancer tumor cells, which can be useful in finding methods to cure, or at least control cancer. Every year, a variety of drugs are being developed to overcome cancer, all of which need to be experimented. Although, these drugs are now experimented on animals, and some human volunteers, the reaction of different patients’ cancer cells to different drugs may differ with the experimented animals, or even with other people. So, there needs to be a method developed to analyze each person’s reaction to the drugs, easily, quickly and without risking the patient’s health. On the other hand, mimicking the in-vivo micro-environment of tumors, the dynamic and continuous perfusion happening in body, is not possible in the already used 2D culture methods. Microfluidic chips are here to solve these issues.In this study, at first, we studied and experimented the static methods used for culturing cancer cells on flask, and also the classic methods of forming spheroids, such as forming them in 96-well dishes, and forming them using hanging droplets in petri dishes. In addition, after the formation of appropriate spheroids, the spheroids were cultured under different concentrations of two different drugs, Cisplatin and Vinorelbine, and then their viability was tested using MTT assay.Next, using the gathered information from the static culture, microfluidics science, and simulation using COMSOL, we designed an integrated microfluidic chip made of PDMS, consisting of two main parts of concentration gradient generator (CGG), and spheroid generator. In the CGG part of this chip, mixtures of medium with 4 different concentrations of drug are generated, each of which lead to a spheroid generator part. In the spheroid generator part, single A549 cancer cells are seeded and trapped in the cylindrical wells, aggregating, and after a while forming spheroids. The final made chip proved to be working well, generating correct concentrations, trapping a large number of the seeded cells, and forming almost spherical spheroids that can be cultured and analyzed under different drug concentrations, and other circumstances, analysis that can be useful for cancer studies
  9. Keywords:
  10. Chips (Circuits) ; Spheroidization ; Tumors ; Concentration Gradient Generator ; Microfluidic Chip

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