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First, surgery itself is associated with increased risk of morbidity and mortality, which may consequently result in delayed/canceled postoperative adjuvant treatment. Therefore, preoperative chemo- or radiotherapy may ensure full dose delivery, unlike for adjuvant treatment. Second, development of metastatic disease during the time frame of neoadjuvant therapy would select patients with advanced systemic disease that might benefit less from more aggressive local regimens including surgery. Third, improved delivery of chemotherapeutic agents may be possible by the still intact blood supply prior to surgery.

With the progress of life time and the increase of the steps, the dilution factor is enhanced. After the 70th passage cell cultures are passed with a dilution factor of 100. To preserve the epithelial cells and fibroblasts, samples are cryopreserved in complete culture medium containing 10% dimethyl sulphoxide in the ratio volume/volume and 30% fetal bovine serum (FBS). This type of solution should always be freshly prepared and not exposed to direct light. The cells to be retained are suspended in 1 mL of this medium for freezing and stored in an ultrafreezer to −80 °C for a few days and then permanently transferred to liquid nitrogen at −196 °C temperature.

13 Laser Microdissection of PDAC Tissue. The figure shows the four most important phases of procedure: (A) Identification of Area; (B) Selection of Areas of interest; (C) Cutting: (D)Harvested fragment of Tissue. Magnification 10×; Hematoxylin staining. 14 Preparation of Primary Cell Culture for LMD. (A) Cells seeding in appropriate chamber and condition to allow the best cell adhesion on PET membrane; original magnification; (B) Primary cell culture on PET membrane. Contrast phase light microscopy, magnification 4×; (C) Magnification 10×.

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