title: Downregulation of H19 improves the differentiation potential of mouse parthenogenetic embryonic stem cells.
abstract: Parthenogenetic embryonic stem cells (P-ESCs) offer an alternative source of pluripotent cells, which hold great promise for autologous transplantation and regenerative medicine. P-ESCs have been successfully derived from blastocysts of several mammalian species. However, compared with biparental embryonic stem cells (B-ESCs), P-ESCs are limited in their ability to fully differentiate into all 3 germ layers. For example, it has been observed that there is a differentiation bias toward ectoderm derivatives at the expense of endoderm and mesoderm derivatives-muscle in particular-in chimeric embryos, teratomas, and embryoid bodies. In the present study we found that H19 expression was highly upregulated in P-ESCs with more than 6-fold overexpression compared with B-ESCs. Thus, we hypothesized that manipulation of the H19 gene in P-ESCs would alleviate their limitations and allow them to function like B-ESCs. To test this hypothesis we employed a small hairpin RNA approach to reduce the amount of H19 transcripts in mouse P-ESCs. We found that downregulation of H19 led to an increase of mesoderm-derived muscle and endoderm in P-ESCs teratomas similar to that observed in B-ESCs teratomas. This phenomenon coincided with upregulation of mesoderm-specific genes such as Myf5, Myf6, and MyoD. Moreover, H19 downregulated P-ESCs differentiated into a higher percentage of beating cardiomyocytes compared with control P-ESCs. Collectively, these results suggest that P-ESCs are amenable to molecular modifications that bring them functionally closer to true ESCs.
authors.affiliation: Department of Animal Science, Michigan State University, East Lansing, MI 48824, USA.
authors.forename: Neli P
authors.initials: NP
authors.lastname: Ragina
authors.forename: Karianne
authors.initials: K
authors.lastname: Schlosser
authors.forename: Jason G
authors.initials: JG
authors.lastname: Knott
authors.forename: Patricia K
authors.initials: PK
authors.lastname: Senagore
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Swiatek
authors.forename: Eun Ah
authors.initials: EA
authors.lastname: Chang
authors.forename: Walid D
authors.initials: WD
authors.lastname: Fakhouri
authors.forename: Brian C
authors.initials: BC
authors.lastname: Schutte
authors.forename: Matti
authors.initials: M
authors.lastname: Kiupel
authors.forename: Jose B
authors.initials: JB
authors.lastname: Cibelli
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D015536:Down-Regulation; D004475:Ectoderm / Q000378:metabolism / Q000473:pathology; D058732:Embryoid Bodies / Q000378:metabolism / Q000502:physiology; D053595:Embryonic Stem Cells / Q000502:physiology* / Q000637:transplantation; D004707:Endoderm / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D005260:Female; D020869:Gene Expression Profiling; D041722:Genes, Transgenic, Suicide; D018392:Genomic Imprinting; D007335:Insulin-Like Growth Factor II / Q000235:genetics / Q000378:metabolism; D059785:Karyotype; D008648:Mesoderm / Q000473:pathology / Q000502:physiology; D051379:Mice; D009132:Muscles / Q000473:pathology; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000502:physiology; D010312:Parthenogenesis; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000235:genetics* / Q000378:metabolism; D013724:Teratoma / Q000378:metabolism / Q000473:pathology
chemical_list: C094335:H19 long non-coding RNA; C513121:IGF2 protein, mouse; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D007335:Insulin-Like Growth Factor II