A1 Refereed original research article in a scientific journal
Expression of leukemia inhibitory factor and its receptors is increased during differentiation of human embryonic stem cells
Authors: Aghajanova L, Skottman H, Strömberg AM, Inzunza J, Lahesmaa R, Hovatta O
Publication year: 2006
Journal: Fertility and Sterility
Journal name in source: Fertility and sterility
Journal acronym: Fertil Steril
Volume: 86
Issue: 4 Suppl
First page : 1193
Last page: 209
Number of pages: 17
ISSN: 0015-0282
eISSN: 1556-5653
DOI: https://doi.org/10.1016/j.fertnstert.2005.12.081
Abstract
To investigate gene expression profiles during the early spontaneous differentiation of human embryonic stem cells (hESCs), with particular emphasis on leukemia inhibitory factor (LIF)-induced pathways and the ultrastructural surface morphology of the undifferentiated and spontaneously differentiated hESCs.\nProspective experimental study.\nUniversity laboratory.\nFour hESC cell lines.\nThe effect of LIF on receptor expression level was studied in cultures.\nGene expression in the hESC line HS237 was analyzed using microarrays. Real-time reverse-transcription polymerase chain reaction was used to validate the microarray results in four hESC lines (HS181, HS235, HS237, HS293). Immunohistochemistry was used to assay LIF, LIF receptor, and gp130 protein expression. Cell surface morphology was studied using scanning electron microscopy.\nThe expression of LIF, LIF receptor, and gp130 messenger RNA and protein was increased in spontaneously differentiated HS237 cells compared with undifferentiated cells, with high expression of an inhibitor of LIF-mediated signaling, suppressor of cytokine signaling-1, in undifferentiated hESCs. Genes, those expressed specifically and those shared in undifferentiated hESCs, differentiated cells, and in fibroblasts, were identified. Supplementation with LIF did not affect the LIF receptor expression.\nThe expression of LIF and its receptors is low in undifferentiated hESCs but increases during differentiation. Added LIF does not prevent spontaneous differentiation. Suppressor of cytokine signaling-1 may prevent LIF signaling in hESCs.\nOBJECTIVE\nDESIGN\nSETTING\nPATIENT(S)\nINTERVENTION(S)\nMAIN OUTCOME MEASURE(S)\nRESULT(S)\nCONCLUSION(S)
To investigate gene expression profiles during the early spontaneous differentiation of human embryonic stem cells (hESCs), with particular emphasis on leukemia inhibitory factor (LIF)-induced pathways and the ultrastructural surface morphology of the undifferentiated and spontaneously differentiated hESCs.\nProspective experimental study.\nUniversity laboratory.\nFour hESC cell lines.\nThe effect of LIF on receptor expression level was studied in cultures.\nGene expression in the hESC line HS237 was analyzed using microarrays. Real-time reverse-transcription polymerase chain reaction was used to validate the microarray results in four hESC lines (HS181, HS235, HS237, HS293). Immunohistochemistry was used to assay LIF, LIF receptor, and gp130 protein expression. Cell surface morphology was studied using scanning electron microscopy.\nThe expression of LIF, LIF receptor, and gp130 messenger RNA and protein was increased in spontaneously differentiated HS237 cells compared with undifferentiated cells, with high expression of an inhibitor of LIF-mediated signaling, suppressor of cytokine signaling-1, in undifferentiated hESCs. Genes, those expressed specifically and those shared in undifferentiated hESCs, differentiated cells, and in fibroblasts, were identified. Supplementation with LIF did not affect the LIF receptor expression.\nThe expression of LIF and its receptors is low in undifferentiated hESCs but increases during differentiation. Added LIF does not prevent spontaneous differentiation. Suppressor of cytokine signaling-1 may prevent LIF signaling in hESCs.\nOBJECTIVE\nDESIGN\nSETTING\nPATIENT(S)\nINTERVENTION(S)\nMAIN OUTCOME MEASURE(S)\nRESULT(S)\nCONCLUSION(S)
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