Boy-Lefèvre M L, Nefussi J R, Paulin D, Hartmann D J, Ricard-Blum S, Herbage D, Forest N
J Histochem Cytochem. 1986 Jul;34(7):835-9. doi: 10.1177/34.7.3519748.
Collagen immunotyping by indirect immunofluorescence was performed in order to investigate the sequential development of bone formation. Osseous tumors were obtained after subcutaneous injection of 3/A/1D-1 teratocarcinoma cell line into 129/Sv mice (Nicolas et al., 1980). Frozen sections of developing tumors were incubated with specific antibodies directed against Types I, II, III, IV, and IX collagens. On Day 9, the expression of Type I and Type III collagens was correlated with the proliferation of mesenchymal cells. From Day 10, chondrogenesis was characterized by the occurrence of cartilaginous collagens, Types II and IX, in the cartilage matrix. Type IV collagen was also detected in focal areas and revealed vascular invasion of the tumor. On Day 13, osteogenesis was demonstrated by the presence of Type I collagen in the bone matrix coating the surfaces. Immunolocalization of Type III collagen on the hemopoietic elements corresponded with the bone remodeling. The sequential transitions of collagen types confirm the development of an endochondral bone tumor. These results suggest that 3/A/1D-1 teratocarcinoma cell line constitutes a valuable system for in vitro study of endochondral bone formation and cell differentiation.
为了研究骨形成的连续发育过程，采用间接免疫荧光法进行胶原蛋白免疫分型。将3/A/1D-1畸胎瘤细胞系皮下注射到129/Sv小鼠体内后获取骨肿瘤（Nicolas等人，1980年）。将发育中肿瘤的冰冻切片与针对I型、II型、III型、IV型和IX型胶原蛋白的特异性抗体一起孵育。在第9天，I型和III型胶原蛋白的表达与间充质细胞的增殖相关。从第10天开始，软骨形成的特征是在软骨基质中出现软骨胶原蛋白II型和IX型。在局部区域也检测到IV型胶原蛋白，显示肿瘤有血管侵袭。在第13天，在覆盖表面的骨基质中存在I型胶原蛋白证明有骨生成。III型胶原蛋白在造血成分上的免疫定位与骨重塑一致。胶原蛋白类型的连续转变证实了软骨内骨肿瘤的发育。这些结果表明，3/A/1D-1畸胎瘤细胞系构成了一个用于软骨内骨形成和细胞分化体外研究的有价值的系统。
