297 DERIVATION AND CHARACTERIZATION OF THE TRANSGENIC SOMATIC CELL NUCLEAR TRANSFER-DERIVED BOVINE EMBRYONIC STEM CELLS
| Autor: | Kyung-Jun Uh, E.-B. Jung, Woo-Suk Hwang, Hyo J. Lee, Hyun-Chang Kim, Sang-Hwan Hyun, Taeyoung Shin, Yeun Wook Kim, S. H. Jeong |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
KOSR
Somatic cell Embryogenesis Cell Biology Embryonic stem cell Cell biology Endocrinology medicine.anatomical_structure Reproductive Medicine Cell culture Immunology Genetics medicine Inner cell mass Somatic cell nuclear transfer Animal Science and Zoology Molecular Biology Developmental Biology Biotechnology |
| Zdroj: | Reproduction, Fertility and Development. 23:246 |
| ISSN: | 1031-3613 |
| DOI: | 10.1071/rdv23n1ab297 |
| Popis: | Bovine transgenic embryonic stem (ES) cells have not been reported yet because it seems that the derivation methods and the culture conditions for the inner cell mass are neither consistent nor optimized. Isolation of inner cell mass and primary culture of ES colonies is a critical step toward the establishment of authentic bovine ES cell lines. Herein, we reconstructed somatic cell nuclear transferred (SCNT) bovine blastocysts carrying a vector expressing the human INF-α gene, and isolated inner cell masses to derive transgenic bovine embryonic stem cells. In addition, we added 2 inhibitors, inhibition (2i system) of the mitogen-activated protein kinase (Erk1/2) cascade, PD0325901(3 Î1/4M), and of glycogen synthase kinase 3, CHIR99021 (1 Î1/4M), in the inner cell mass primary culture to check reliability of the 2i system for bovine ES culture. The 2 inhibitors made the morphology of colonies more intact, and primary colonies were better maintained in early passages. However, there were no significant effects on the attachment rate and maintenance in late passages (percent of percent over 3 passages: 2i system, 21/38 (55.3%); control, 22/42 (33.3%); P |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|