Figure 2

Macroscopic examination of the CAM and implanted human NCI-H446 cells. The entire experimental process from the implantation of NCI-H446 cells on the CAM and the formation of the transplantation tumor is shown. (A) Irregular window made in the egg shell of a 7-day-old chick embryo. (B) Elimination of the chick embryo in the CAM was observed. (C) The CAM was peeled for the assay. (D) Diagram of the technique for the implantation of NCI-H446 cells onto the CAM. (E) Diagram of the technique for the formation of the transplantation tumor. (F) The transplantation tumor (white mass was pointed by the tip) was formed on the side facing the chick embryo. (G-H) Histological evaluation of the transplanted tumor on the CAM by hematoxylin-eosin staining is shown:(G) The structure of the transplantation tumor and peripheral vessels (50 ×). (H) Pathological appearance of the transplantation tumor (200 ×). (I) Specific analysis was carried out by immunohistochemistry for the expression of NSE. The cellular nucleus was irregular, and positive expression for NSE was found in the intercellular substance or endochylema (400 ×).