Biomedica : revista del Instituto Nacional de Salud · 2006
[Immunocytochemical and molecular studies with primary cultures of molar tissue].
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Bernal Yinth Andrea, Díaz Luis Eduardo, Acosta Jinneth, Crane Cecilia, Carrasco-Rodríguez Stella, Bermúdez Antonio José, Sánchez-Gómez Myriam
Аннотация
INTRODUCTION: Gestational trophoblastic disease includes a group of pathologies characterized by abnormal trophoblast growth and invasion. The molecular bases of the disease are largely unknown, due in part to the lack of appropriate biological models. The insulin-like growth factor (IGF) system plays a fundamental role in the growth and development of many tissues and is involved in the progression of several diseases.
OBJECTIVES: Primary cell cultures derived from first trimester placenta were characterized from patients with complete hydatidiform mole and spontaneous non molar abortion by immunocytochemical and molecular methods.
MATERIALS AND METHODS: The immunocytochemical method used specific markers for trophoblastic cells, whereas RT-PCR was used to identify insulin-like growth factor gene expression.
RESULTS: Histochemical staining with hematoxilin-eosin revealed that the cultures contained heterogeneous cell types, including trophoblast and endometrial decidual cells. The ratio of trophoblast cells in the cultures varied between 16% and 37%, as detected by cytokeratine-7 as the specific trophoblast marker. Gene expression analysis corroborated the presence of trophoblasts by detecting insulin-like growth factor II mRNA, whereas GH-V transcripts were correlated with the presence of syncitiotrophoblasts. Insulin-like growth factor I and insulin-like growth factor binding protein 1 mRNAs were related to mesenchyimal and decidual cells, respectively. Higher insulin-like growth factor II expression levels were found in molar tissues in comparison with non-molar abortions.
CONCLUSION: By combining three methodologies-morphology, immunocytochemistry and gene expression, characterization and follow-up of placenta cultures from abnormal tissues is found to facilitate diagnosis.
OBJECTIVES: Primary cell cultures derived from first trimester placenta were characterized from patients with complete hydatidiform mole and spontaneous non molar abortion by immunocytochemical and molecular methods.
MATERIALS AND METHODS: The immunocytochemical method used specific markers for trophoblastic cells, whereas RT-PCR was used to identify insulin-like growth factor gene expression.
RESULTS: Histochemical staining with hematoxilin-eosin revealed that the cultures contained heterogeneous cell types, including trophoblast and endometrial decidual cells. The ratio of trophoblast cells in the cultures varied between 16% and 37%, as detected by cytokeratine-7 as the specific trophoblast marker. Gene expression analysis corroborated the presence of trophoblasts by detecting insulin-like growth factor II mRNA, whereas GH-V transcripts were correlated with the presence of syncitiotrophoblasts. Insulin-like growth factor I and insulin-like growth factor binding protein 1 mRNAs were related to mesenchyimal and decidual cells, respectively. Higher insulin-like growth factor II expression levels were found in molar tissues in comparison with non-molar abortions.
CONCLUSION: By combining three methodologies-morphology, immunocytochemistry and gene expression, characterization and follow-up of placenta cultures from abnormal tissues is found to facilitate diagnosis.