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Human pluripotent embryonal carcinoma NTERA2 cl.D1 cells maintain their typical morphology in an angiomyogenic medium.

Simões PD, Ramos T - J Negat Results Biomed (2007)

Bottom Line: Among them were early and late cardiac, endothelial, neuronal and teratocarcinoma genes.Our results suggest that despite the NT2/D1 cells natural tendency to differentiate into neuroectodermal lineages, they can activate genes of mesodermal lineages.Therefore, we believe that these pluripotent cells might still be a good model to study biological development of mesodermal derivatives, provided the right culture conditions are met.

View Article: PubMed Central - HTML - PubMed

Affiliation: Instituto de Tecnologia Biomédica, Laboratório de Biomateriais, Faculdade de Medicina Dentária da Universidade de Lisboa, Cidade Universitária, Lisbon, Portugal. pedrodanielsim@yahoo.co.uk

ABSTRACT

Background: Pluripotent embryonal carcinomas are good potential models, to study, "in vitro," the mechanisms that control differentiation during embryogenesis. The NTERA2cl.D1 (NT2/D1) cell line is a well known system of ectodermal differentiation. Retinoic acid (RA) induces a dorsal pattern of differentiation (essentially neurons) and bone morphogenetic protein (BMP) or hexamethylenebisacetamide (HMBA) induces a more ventral (epidermal) pattern of differentiation. However, whether these human cells could give rise to mesoderm derivatives as their counterpart in mouse remained elusive. We analyzed the morphological characteristics and transcriptional activation of genes pertinent in cardiac muscle and endothelium differentiation, during the growth of NT2/D1 cells in an inductive angiomyogenic medium with or without Bone Morphogenetic Protein 2 (BMP2).

Results: Our experiments showed that NT2/D1 maintains their typical actin organization in angiomyogenic medium. Although the beta myosin heavy chain gene was never detected, all the other 15 genes analyzed maintained their expression throughout the time course of the experiment. Among them were early and late cardiac, endothelial, neuronal and teratocarcinoma genes.

Conclusion: Our results suggest that despite the NT2/D1 cells natural tendency to differentiate into neuroectodermal lineages, they can activate genes of mesodermal lineages. Therefore, we believe that these pluripotent cells might still be a good model to study biological development of mesodermal derivatives, provided the right culture conditions are met.

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NT2/D1 cells in expansion medium. Phase contrast images of three typical morphologies of NT2/D1 cells in expansion medium, (RPMI, FBS and antibiotics) analyzed in a CK2 Olympus microscope: A. Embryoid body-like structures, magnification 40×; B. epithelioid-like phenotype, magnification 100×; C. neuronal-like phenotype, magnification 200×.
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Figure 1: NT2/D1 cells in expansion medium. Phase contrast images of three typical morphologies of NT2/D1 cells in expansion medium, (RPMI, FBS and antibiotics) analyzed in a CK2 Olympus microscope: A. Embryoid body-like structures, magnification 40×; B. epithelioid-like phenotype, magnification 100×; C. neuronal-like phenotype, magnification 200×.

Mentions: The typical NT2/D1 morphological characteristics, as the increase in nuclear/cytoplasm volume ratio, the flattening of the cells, the prominent nucleoli (Figure 1), the polymerized actin organization (phalloidin staining) and ultrastructural analysis were used as the morphological criteria to monitor cell differentiation.


Human pluripotent embryonal carcinoma NTERA2 cl.D1 cells maintain their typical morphology in an angiomyogenic medium.

Simões PD, Ramos T - J Negat Results Biomed (2007)

NT2/D1 cells in expansion medium. Phase contrast images of three typical morphologies of NT2/D1 cells in expansion medium, (RPMI, FBS and antibiotics) analyzed in a CK2 Olympus microscope: A. Embryoid body-like structures, magnification 40×; B. epithelioid-like phenotype, magnification 100×; C. neuronal-like phenotype, magnification 200×.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1863432&req=5

Figure 1: NT2/D1 cells in expansion medium. Phase contrast images of three typical morphologies of NT2/D1 cells in expansion medium, (RPMI, FBS and antibiotics) analyzed in a CK2 Olympus microscope: A. Embryoid body-like structures, magnification 40×; B. epithelioid-like phenotype, magnification 100×; C. neuronal-like phenotype, magnification 200×.
Mentions: The typical NT2/D1 morphological characteristics, as the increase in nuclear/cytoplasm volume ratio, the flattening of the cells, the prominent nucleoli (Figure 1), the polymerized actin organization (phalloidin staining) and ultrastructural analysis were used as the morphological criteria to monitor cell differentiation.

Bottom Line: Among them were early and late cardiac, endothelial, neuronal and teratocarcinoma genes.Our results suggest that despite the NT2/D1 cells natural tendency to differentiate into neuroectodermal lineages, they can activate genes of mesodermal lineages.Therefore, we believe that these pluripotent cells might still be a good model to study biological development of mesodermal derivatives, provided the right culture conditions are met.

View Article: PubMed Central - HTML - PubMed

Affiliation: Instituto de Tecnologia Biomédica, Laboratório de Biomateriais, Faculdade de Medicina Dentária da Universidade de Lisboa, Cidade Universitária, Lisbon, Portugal. pedrodanielsim@yahoo.co.uk

ABSTRACT

Background: Pluripotent embryonal carcinomas are good potential models, to study, "in vitro," the mechanisms that control differentiation during embryogenesis. The NTERA2cl.D1 (NT2/D1) cell line is a well known system of ectodermal differentiation. Retinoic acid (RA) induces a dorsal pattern of differentiation (essentially neurons) and bone morphogenetic protein (BMP) or hexamethylenebisacetamide (HMBA) induces a more ventral (epidermal) pattern of differentiation. However, whether these human cells could give rise to mesoderm derivatives as their counterpart in mouse remained elusive. We analyzed the morphological characteristics and transcriptional activation of genes pertinent in cardiac muscle and endothelium differentiation, during the growth of NT2/D1 cells in an inductive angiomyogenic medium with or without Bone Morphogenetic Protein 2 (BMP2).

Results: Our experiments showed that NT2/D1 maintains their typical actin organization in angiomyogenic medium. Although the beta myosin heavy chain gene was never detected, all the other 15 genes analyzed maintained their expression throughout the time course of the experiment. Among them were early and late cardiac, endothelial, neuronal and teratocarcinoma genes.

Conclusion: Our results suggest that despite the NT2/D1 cells natural tendency to differentiate into neuroectodermal lineages, they can activate genes of mesodermal lineages. Therefore, we believe that these pluripotent cells might still be a good model to study biological development of mesodermal derivatives, provided the right culture conditions are met.

Show MeSH
Related in: MedlinePlus