Human embryonic germ cells for future neuronal replacement therapy

Turnpenny, Lee; Cameron, Iain; Spalluto, Cosma; Hanley, Karen Piper; Wilson, David I.; Hanley, Neil A.

doi:10.1016/j.brainresbull.2005.08.014

Cited by 19 publications

(14 citation statements)

References 37 publications

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“…Furthermore, these cells can proliferate indefinitely without senescence [3,6], allowing the production of large amounts of biological material. These properties make ESCs and EGCs very attractive candidates for cell replacement therapy in various degenerative diseases that are associated with loss of functional cells, such as type I diabetes or Alzheimer diseases [9,10]. They can also have applications for in vitro studies of embryogenesis, abnormal development, gene discovery, drug and teratogen testing, as well as for understanding genetic syndromes [11,12].…”

Section: Introductionmentioning

confidence: 97%

Proteome analysis of the culture environment supporting undifferentiated mouse embryonic stem and germ cell growth

et al. 2007

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The therapeutical interest of pluripotent cells and ethical issues related to the establishment of human embryonic stem cell (ESC) or embryonic germ cell (EGC) lines raise the understanding of the mechanism underlying pluripotency to a fundamental issue. Establishing a protein pluripotency signature for these cells can be complicated by the presence of unrelated proteins produced by the culture environment. Here, we have analyzed the environment supporting ESC and EGC growth, and established 2-D reference maps for each constituent present in this culture environment: mouse embryonic fibroblast feeder cells, culture medium (CM) and gelatin. The establishment of these reference maps is essential prior to the study of ESC and EGC specific proteomes. Indeed, these maps can be subtracted from ESC or EGC maps to allow focusing on spots specific for ESCs or EGCs. Our study led to the identification of 110 unique proteins from fibroblast feeder cells and 23 unique proteins from the CM, which represent major contaminants of ESC and EGC proteomes. For gelatin, no collagen-specific proteins were identified, most likely due to difficulties in resolution and low quantities. Furthermore, no differences were observed between naive and conditioned CM. Finally, we compared these reference maps to ESC 2-D gels and isolated 17 ESC specific spots. Among these spots, proteins that had already been identified in previous human and mouse ESC proteomes were identified but no apparent ESC-specific pluripotency marker could be identified. This work represents an essential step in furthering the knowledge of environmental factors supporting ESC and EGC growth.

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Section: Introductionmentioning

confidence: 97%

Proteome analysis of the culture environment supporting undifferentiated mouse embryonic stem and germ cell growth

et al. 2007

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“…ESCs and EGCs thus provide exciting models for understanding the underlying mechanisms that make a cell pluripotent [1][2][3]. ESCs and EGCs are not only valuable research tools in cell biology, but they also bring promise for cell-based therapies [4][5][6][7]. These therapies are based on the production of specific cell types in order to restore, after transplantation, physiological functions in degenerative diseases such as diabetes or Parkinson diseases.…”

Section: Introductionmentioning

confidence: 97%

Nuclear proteome analysis of undifferentiated mouse embryonic stem and germ cells

et al. 2008

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Embryonic stem cells (ESCs) and embryonic germ cells (EGCs) provide exciting models for understanding the underlying mechanisms that make a cell pluripotent. Indeed, such understanding would enable dedifferentiation and reprogrammation of any cell type from a patient needing a cell therapy treatment. Proteome analysis has emerged as an important technology for deciphering these biological processes and thereby ESC and EGC proteomes are increasingly studied. Nevertheless, their nuclear proteomes have only been poorly investigated up to now. In order to investigate signaling pathways potentially involved in pluripotency, proteomic analyses have been performed on mouse ESC and EGC nuclear proteins. Nuclei from ESCs and EGCs at undifferentiated stage were purified by subcellular fractionation. After 2-D separation, a subtractive strategy (subtracting culture environment contaminating spots) was applied and a comparison of ESC, (8.5 day post coïtum (dpc))-EGC and (11.5 dpc)-EGC specific nuclear proteomes was performed. A total of 33 ESC, 53 (8.5 dpc)-EGC, and 36 (11.5 dpc)-EGC spots were identified by MALDI-TOF-MS and/or nano-LC-MS/MS. This approach led to the identification of two isoforms (with and without N-terminal acetylation) of a known pluripotency marker, namely developmental pluripotency associated 5 (DPPA5), which has never been identified before in 2-D gel-MS studies of ESCs and EGCs. Furthermore, we demonstrated the efficiency of our subtracting strategy, in association with a nuclear subfractionation by the identification of a new protein (protein arginine N-methyltransferase 7; PRMT7) behaving as proteins involved in pluripotency.

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“…The process of neural differentiation accompanied with neuroepithelial-like cells (NELC) generation which are multipotent stem cells that give rise to all the cells of central nervous system, including various types of neurons, astrocytes, and oligodendrocytes (Turnpenny et al 2005). Today, different sources and methods are currently used for derivation neuroepithelial-like cells in vitro (Silani et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Alteration in genes expression patterns during in vitro differentiation of mouse spermatogonial cells into neuroepithelial-like cells

et al. 2012

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Pluripotent stem cells derived from testis is a new, natural, and unlimited source for cell therapy in regenerative medicine and represent a possible alternative to replacing of all cells in the body. Here, we designed a simple co-culture system of spermatogonia cells with Sertoli cells for the generation of embryonic stem-like cells from mouse testis. The importance of our simple method will be clear when we compared it with other complex and time-consuming methods. Embryonic stem-like colonies with sharp border confirmed by real-time PCR, immunocytochemistry and flow cytometry assessments. Embryonic stem-like colonies were immunopositive for pluripotency markers. Transition of spermatogonia cells to embryonic stem-like cells was accompanied by extensive changes in gene expression. These changes included significant increase in pluripotency genes expression and significant decrease in germ cellspecific genes expression. Also, we proved the differentiation capacity of embryonic stem-like cells to neuroepithelial-like cells which were immunoreactive to Nestin and Neurofilament 68. Evaluation of genes expression during in vitro differentiation into neuroepithelial-like cells showed high-level expression of Nestin whether this gene approximately has no expression in undifferentiated embryonic stem-like cells. Also, expression of pluripotency genes has significantly decreased in neuroepithelial-like cells compared with embryonic stem-like cells. This study shows that embryonic stem-like cells derived from testis are capable to differentiate into neuroepitheliallike cells that may provide a cellular reservoir usable for neurodegenerative disorders.

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Human embryonic germ cells for future neuronal replacement therapy

Cited by 19 publications

References 37 publications

Proteome analysis of the culture environment supporting undifferentiated mouse embryonic stem and germ cell growth

Proteome analysis of the culture environment supporting undifferentiated mouse embryonic stem and germ cell growth

Nuclear proteome analysis of undifferentiated mouse embryonic stem and germ cells

Alteration in genes expression patterns during in vitro differentiation of mouse spermatogonial cells into neuroepithelial-like cells

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