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Title: SPONTANEOUS DIFFERENTIATION OF PORCINE AND BOVINE EMBRYONIC STEM CELLS (EPIBLAST) INTO ASTROCYTES OR NEURONS.

Author
item Talbot, Neil
item Powell, Anne
item Garrett, Wesley

Submitted to: In Vitro Cellular And Developmental Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/25/2002
Publication Date: 7/5/2002
Citation: Talbot, N.C., Powell, A.M., Garrett, W.M. Spontaneous differentiation of porcine and bovine embryonic stem cells epiblast) into astrocytes or neurons. In Vitro Cellular And Developmental Biology. 38:191-197. 2002.

Interpretive Summary: True ES cells are theoretically capable of changing ("differentiating") into all the cell types found in the body. The culture of putative bovine and porcine embryonic stem (ES) cells resulted in spontaneous differentiation into cells resembling nervous tissue cells. An analysis of the cells was undertaken to assess whether these cells had "markers" or proteins in them that are typical of nerve cells or nerve accessory cells. The present study indicates that these bovine and porcine ES- cell-derived cell cultures are in fact cultures of nerves or nervous tissue cells that support the nerves. Thus, the study confirms the ES- like nature of the bovine and porcine ES cell cultures and defines a source of potential nerve stem cell cultures. These findings suggest that these cells may be an excellent resource for scientists conducing research on nuclear transfer.

Technical Abstract: The immunocytochemical characterization of bovine and porcine epiblast- derived cell cultures that morphologically appeared to be derivatives of neuroectoderm, i.e., "neuron-like" cells, is reported. From among several neuron-like cell culture isolated, two porcine (PICM-10 and PICM-11) and one bovine (CICM-3) were examined. The morphology of PICM and CICM neuron-like cell cultures varied somewhat but presented as fibroblastic to dendritic as observed by phase-contrast microscopy. The cells were reacted with rabbit polyclonal anti-GFAP and mouse monoclonals specific for vimentin, alpha-internexin, tyrosine hydroxylase, or pan-axonal neurofilament. STO feeder cells were not reactive with any of the antibodies tested. The PICM-10 and -11 were positive for reactivity with anti-vimentin and anti-GFAP antibodies, but were negative for neurofilament, alpha-internexin, and tyrosine hydroxylase antibody reactivity. Most or all CICM-3 cells were positive for reaction with anti-vimentin antibodies and a small percentage of the population (less than 1%) immunostained positively for alpha-internexin and neurofilaments. The data are consistent with the possibility that the CICM-3 cells may be a neuronal precursor cells and that PICM-10 and 11 cells are cultures of Type 2 astrocytes. The study proves that the spontaneous in vitro differentiation potential of porcine and bovine epiblast cells includes cells of neuroectodermal origin.