Immunocytochemical and molecular studies with primary cultures of molar tissue
Keywords:
Hydatidiform mole, invasive, insulin-like growth factor I, insulin-like growth factor II, insulin-like growth factor binding protein 1
Abstract
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.
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References
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11. Han VK, Carter AM. Spatial and temporal patterns of expression of messenger RNA for Insulin-like growth factors and their binding proteins in the placenta of man and laboratory animals. Placenta 2000;21:289-305.
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14. Pötgens AJ, Kataokab H, Ferstla S, Frank HG, Kaufmanna P. A positive immunoselection method to isolate villous cytotrophoblast cells from first trimester and term placenta to high purity. Placenta 2003;24:412-23.
15. Irving JA, Lysiak JJ, Graham CH, Hearn S, Han VK, Lala PK. Characteristics of trophoblast cells migrating from first trimester chorionic villus explants and propagated in culture. Placenta 1995;16:413-33.
2. Giudice LC, Irwin JC. Roles of the insulin-like growth factor family in nonpregant human endometrium and at the decidual:trophoblast interface. Semin Reprod Endocrinol 1999;17:13-21.
3. Ezpeleta J, López A. Enfermedad trofoblástica gestacional. Rev Esp Patol 2002;35:187-200.
4. Balaram P, John M, Enose J, Symaladevi PK. Demonstration of TGF-á-EGFR and EGF-EGFR autocrine loops and their relation to proliferation in complete hydatidiform moles (CHM). Int J Gynecol Cancer 2001;11:397-402.
5. Wihman IL, Carlstrom K, Faxen M. Insulin-like growth factor-I in women with hydatidiform mole and in normal pregnancy. Obstet Gynecol 1998;92:431-4.
6. Han VK, Basset N, Walton J, Challis JR. The expression of insulin-like growth factor (IGF) and IGF-binding protein (IGFBP) genes in the human placenta and membranes: Evidence for IGF-IGFBP interactions at the feto-maternal interfase. J Clin Endocrinol Metab 1996;81:2680-93.
7. Lacey H, Haigh T, Westwood M, Aplin JD. Mesenchymally-derived insulin-like growth provides a paracrine stimulus for trophoblast migration. BMC Dev Biol 2002; 2:5.
8. Nayak NR, Guidice LC. Comparative biology of the IGF system in endometrium, decidua, and placenta, and clinical implications for foetal growth and implatantion disorders. Placenta 2003;24:281-96.
9. Yagel S, Casper RF, Powell W, Parhar RS, Lala PK. Characterization of pure human first-trimester cytotrophoblast cells in long-term culture: Growth pattern, markers and hormone production. Am J Obstet Gynecol 1989;160:938-45.
10. Chen JC, Ornelles DA, Shenk T. The adenovirus l3 23-kilodalton proteinase cleaves the amino-terminal head domain from cytokeratin 18 and disrupts the cytokeratin network of HeLa cells. J Virol 1993;67:3507-14.
11. Han VK, Carter AM. Spatial and temporal patterns of expression of messenger RNA for Insulin-like growth factors and their binding proteins in the placenta of man and laboratory animals. Placenta 2000;21:289-305.
12. Blaschitz A, Weiss U, Dohr G, Desoye G. Antibody reaction patterns in first trimester placenta: implications for trophoblast isolation and purity screening. Placenta 2000;21:733-41.
13. Tarrade A, Kuen RL, Malassiné A, Tricottet V, Blain P, Vidaud M, et al. Characterization of human villous and extravillous trophoblast isolated from first trimester placenta. Lab Invest 2001;81:1199-211.
14. Pötgens AJ, Kataokab H, Ferstla S, Frank HG, Kaufmanna P. A positive immunoselection method to isolate villous cytotrophoblast cells from first trimester and term placenta to high purity. Placenta 2003;24:412-23.
15. Irving JA, Lysiak JJ, Graham CH, Hearn S, Han VK, Lala PK. Characteristics of trophoblast cells migrating from first trimester chorionic villus explants and propagated in culture. Placenta 1995;16:413-33.
How to Cite
1.
Bernal YA, Díaz LE, Acosta J, Crane C, Carrasco-Rodríguez S, Bermúdez AJ, et al. Immunocytochemical and molecular studies with primary cultures of molar tissue. biomedica [Internet]. 2006 Dec. 1 [cited 2024 May 11];26(4):509-16. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/316
Published
2006-12-01
Issue
Section
Original articles
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