细胞注射法及组织块移植法在构建异位妊娠动物模型比较

Comparision between Cell Suspension Inoculation and Tissue Block Transplantation in Establishment of Ectopic Pregnancy Animal Models

目的：比较滋养细胞悬液注射法与绒毛组织块移植法在异位妊娠动物模型中的可行性。方法：收集新鲜宫内妊娠绒毛标本30例,其中20例培养成滋养层细胞,并将传代细胞分别植入BALB/c裸鼠腹部皮下（A组）、腹壁下（B组）、宫角（C组）;另外10例绒毛组织块分别植入裸鼠盆腔内（D组）,每组6只。观察种植后第7天、第15天肿块大小的变化;第7天,各组随机选取2只成瘤裸鼠处死,解剖、切取瘤体,形态学检测并免疫组化检测;第15天处死全部裸鼠,解剖、切取瘤体,形态学检测并免疫组化检测。结果：第7天：（1）瘤体观察：A组有2只裸鼠未见瘤体形成;B组有1只未见瘤体形成;C组全部未见瘤体形成;D组全部见瘤体,组织块瘤体与机体部分融合。（2）形态学检测：A组：皮下囊肿,包膜完整,内部可见移植的细胞;B组：皮下可见移植的细胞;C组：未见种植细胞;D组：边缘绒毛滋养细胞细胞核清晰。（3）免疫组化：A组：CK18在滋养细胞胞浆呈阳性表达,滋养细胞细胞核不清或消失;B组：CK18在滋养细胞胞浆中呈阳性表达,滋养细胞细胞核不清或消失;C组：未见种植细胞;D组：CK18在滋养细胞胞浆呈阳性表达,边缘绒毛滋养细胞细胞核清晰。第15天：（1）瘤体观察：A组：有2只分别在第8天、第10天死亡,剩余2只,未见瘤体形成;B组：有1只在第8天死亡,剩余3只,有1只未见瘤体形成,另2只瘤体较前增大;C组：剩余4只,全部未见瘤体形成;D组：剩余4只,全部见瘤体,组织块瘤体增大明显;（2）形态学检测：A组：未见移植的细胞;B组：皮下可见移植的细胞;C组：未见种植细胞;D组：边缘绒毛滋养细胞细胞核清晰;（3）免疫组化：A组：未见种植细胞;B组：CK18在滋养细胞胞浆中呈阳性表达,滋养细胞细胞核不清或消失;C组：未见种植细胞;D组：CK18在滋养细胞胞浆呈阳性表达,边缘绒毛滋养细胞细胞核清晰。结论：细胞注射法难以建立可靠、稳定的异位妊娠动物模型;组织块移植法可以建立较可靠的动物模型。

Objective：To compare the feasibility of establishment of ectopic pregnancy animal models by cell suspension inoculation and tissue block transplantation.Methods：30 cases of intrauterine pregnancy fresh villus samples were collected.20 cases were cultured into trophoblast cells,and cells were implanted into nude mice subcutaneous（group A）,abdominal wall（group B）,uterine horn（group C）,another 10 cases of villous tissue block were implanted into the pelvic cavity（group D）,and 6 nude mice in each group.At day 7 and 15,the changes of tumor size were observed.At day 7,each group were randomly selected 2 nude mice were sacrificed,dissected and cut the tumor,and examined by morphology and immunohistochemical detection.At day 15,all the mice were sacrificed,dissected and cut the tumor,and examined by morphology and immunohistochemical detection.Results：At day 7：（1）Changes of tumor size：Group A：No tumor formation was found in 2 nude mice.Group B：No tumor formation was found in 1 nude mice.Group C：No tumor formation was observed in all nude mice.Group D：Tumor formation was observed in all nude mice,and the tumor had merged together with the body.（2）Morphology and detection：Group A：subcutaneous cyst,complete capsule,the transplanted cells were visible inside.Group B：The transplanted cells were visible under the subcutaneous tissue.Group C：No transplanted cells were found.Group D：The nuclei of trophoblast cells at the margin of villus were clear.（3）Immunohistochemical detection：Group A：CK18 showed positive expression in the cytoplasm of trophoblast cells,and the nuclei of trophoblast cells were not clear or disappeared.Group B：CK18 showed positive expression in the cytoplasm of trophoblast cells,and the nuclei of trophoblast cells were not clear or disappeared.Group C：No transplanted cells were found.Group D：CK18 showed positive expression in the cytoplasm of trophoblast cells,and the nuclei of trophoblast cells at the margin of villus were clear.At day 15：（1）Changes of tumor size：Group A：There were 2 deaths in eighth days and tenth days,and the remaining 2 showed no tumor formation.Group B：There were 1 deaths in eighth days,and of the remaining 3,1 showed no tumor formation,and the other 2 tumors were larger than before.Group C：No tumor formation was observed in all the remaining 4.Group D：Tumor formation was observed in all the remaining 4,and the tumors were obviously larger than before.（2）Morphology and detection：Group A：No transplanted cells were found.Group B：The transplanted cells were visible under the subcutaneous tissue.Group C：No transplanted cells were found.Group D：The nuclei of trophoblast cells at the margin of villus were clear.（3）Immunohistochemical detection：Group A：No transplanted cells were found.Group B：CK18 showed positive expression in the cytoplasm of trophoblast cells,and the nuclei of trophoblast cells were not clear or disappeared.Group C：No transplanted cells were found.Group D：CK18 showed positive expression in the cytoplasm of trophoblast cells,and the nuclei of trophoblast cells at the margin of villus were clear.Conclusion：Cell suspension inoculation is not appropriate to establish ectopic pregnancy animal models.In contrast,the tissue block transplantation is a good choice for establishing reliable ectopic pregnancy animal models.