Embryogenesis and blastocyst development after somatic cell nuclear transfer in nonhuman primates: overcoming defects caused by meiotic spindle extraction
| Autor: | Saverio Capuano, Tanja Dominko, Christopher S. Navara, Gerald Schatten, Kowit Yu Chong, Sung Keun Kang, Calvin Simerly, Woo Suk Hwang, Duane A. Compton, Byeong Chun Lee, Sang-Hwan Hyun, Gabriella G. Gosman |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Nuclear Transfer Techniques
Paclitaxel Cloning Organism Embryonic Development Kinesins Stem cells Biology Microtubules Cytoplast Chromosomes medicine Animals Humans Blastocyst Molecular Biology Cells Cultured Cytoskeleton Cloning Genetics Molecular Motor Proteins Meiotic spindle extraction Nuclear Proteins Molecular motors Embryo Cell Biology Embryo Transfer Antineoplastic Agents Phytogenic Embryonic stem cell Embryo transfer Cell biology Meiosis medicine.anatomical_structure NuMA Fertilization Embryogenesis Oocytes Macaca Somatic cell nuclear transfer Cattle Female Stem cell Nuclear transfer Developmental Biology |
| Zdroj: | Developmental Biology. 276(2):237-252 |
| ISSN: | 0012-1606 |
| DOI: | 10.1016/j.ydbio.2004.10.006 |
| Popis: | Therapeutic cloning or nuclear transfer for stem cells (NTSC) seeks to overcome immune rejection through the development of embryonic stem cells (ES cells) derived from cloned blastocysts. The successful derivation of a human embryonic stem cell (hESC) line from blastocysts generated by somatic cell nuclear transfer (SCNT) provides proof-of-principle for “therapeutic cloning,” though immune matching of the differentiated NT-hES remains to be established. Here, in nonhuman primates (NHPs; rhesus and cynomologus macaques), the strategies used with human SCNT improve NHP-SCNT development significantly. Protocol improvements include the following: enucleation just prior to metaphase-II arrest; extrusion rather than extraction of the meiotic spindle-chromosome complex (SCC); nuclear transfer by electrofusion with simultaneous cytoplast activation; and sequential media. Embryo transfers (ET) of 135 SCNT-NHP into 25 staged surrogates did not result in convincing evidence of pregnancies after 30 days post-ET. These results demonstrate that (i) protocols optimized in humans generate preimplantation embryos in nonhuman primates; (ii) some, though perhaps not yet all, hurdles in deriving NT-nhpES cells from cloned macaque embryos (therapeutic cloning) have been overcome; (iii) reproductive cloning with SCNT-NHP embryos appears significantly less efficient than with fertilized embryos; (iv) therapeutic cloning with matured metaphase-II oocytes, aged oocytes, or “fertilization failures” might remain difficult since enucleation is optimally performed prior to metaphase-II arrest; and (v) challenges remain for producing reproductive successes since NT embryos appear inferior to fertilized ones due to spindle defects resulting from centrosome and motor deficiencies that produce aneuploid preimplantation embryos, among other anomalies including genomic imprinting, mitochondrial and cytoplasmic heterogeneities, cell cycle asynchronies, and improper nuclear reprogramming. |
| Databáze: | OpenAIRE |
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