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IFMSA-Zaragoza Project

Cell culture involves maintaining cell lines under controlled conditions that differ from their natural environment, focusing on their proliferation and differentiation capabilities. This project highlights essential factors including sterilization, nutrient media, and daily care routines for cultured cells, particularly in animal studies using nude mice. It covers experimental design for intra-tumor injections and measures tumor size to assess treatment efficacy. The study reveals insights into breast tumors derived from nude mice, emphasizing the significance of environmental control in cell culture.

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IFMSA-Zaragoza Project

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Presentation Transcript

  1. El Pilar IFMSA-Zaragoza Project B9602015Chia-Chen Li

  2. Cell Culture

  3. What is CELL CULTURE?! (1) • Maintaining cell line survival under controlled conditions, in despite of their original environment • Proliferative capacity • Differentiation ability • Corresponding cell age

  4. What is CELL CULTURE?! (2) • Controlled conditions • ex vivo • in vitro • Tissue source • Multi-cellular eukaryotes • Mainly animals • Also plants, fungi & microbes

  5. General Requirements (1) • Sterilization • Methanol • Filtering by 0.20 μm • Culture surfaces • Petri dish, flake or plate • Incubation • Temperature at 37℃

  6. General Requirements (2) • Nutrient media • Antibiotic • Buffering • efficient at pH 7.2-7.6 • CO2/pH detectable • Red (pH 8, fresh) Yellow (pH 6, wastes) • Trypsin, the growth inhibitor

  7. Daily Care • Changing media (yellow) • Microscopy observation for cell growth monitoring, monolayer • Trypsin • Reduction for multilayer culture • Dividing for new flakes • Photographing

  8. Results (1) • Breast tumor • derived from nude mice • Hyperchromatic • Hyper-proliferative

  9. Prominent nuclei

  10. Results (2) • Breast tumor • from nude mice • Duct • Ductules ductules duct

  11. Results (3) • Breast tumor • derived from nude mice • Epithelial cell alignment

  12. cell alignment

  13. Animal Experiment

  14. Athymic Nude Mice • Spontaneous genetic mutated mice • FOXN1 deletion • Immunodeficient • Lack of mature T cell • Lack of adaptive immune response • Xenograft transplant • Human prostate tumor

  15. Woking Conditions • Requirements • Working robe • Gloves • Shoe pads • Not allowed • Bare-handed touching • Sniffing & etc.

  16. Daily Care for Nude Mice • Provides • Meal, sterilized • Water, sterilized • Bed surface • Used cages must be washed and sterilized for another-time use.

  17. Experiment Design • Intra-tumor injection, 5 times / 3 weeks • Control • normal saline buffer • Variables • Anti-proliferative ligands • Granulysin • Tumor size measuring • Every injection • The middle of each injection intervals Under anaesthesia

  18. Efficiency Determination • By square a*b, the 2D area of tumor • Effective potency: • The effectiveness can be confirmed if there’s significant change of the potency, in which the tumor size reduces. Experiment is still in progress. a b a1*b1 - a0*b0 a0*b0

  19. Thanks! La Seo

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