Contribution of Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells to Chimeras through Injection and Coculture of Embryos

Abstract

Blastocyst injection and morula aggregation are commonly used to evaluate stem cell pluripotency based on chimeric contribution of the stem cells. To assess the protocols for generating chimeras from stem cells, 8-cell mouse embryos were either injected or cocultured with mouse embryonic stem cells and induced pluripotent stem cells, respectively. Although a significantly higher chimera rate resulted from blastocyst injection, the highest germline contribution resulted from injection of 8-cell embryos with embryonic stem cells. The fully agouti colored chimeras were generated from both injection and coculture of 8-cell embryos with embryonic stem cells. Additionally, microsatellite DNA screening showed that the fully agouti colored chimeras were fully embryonic stem cell derived mice. Unlike embryonic stem cells, the mouse chimeras were only generated from injection of 8-cell embryos with induced pluripotent stem cells and none of these showed germline transmission. The results indicated that injection of 8-cell embryos is the most efficient method for assessing stem cell pluripotency and generating induced pluripotent stem cell chimeras, embryonic stem cell chimeras with germline transmission, and fully mouse embryonic stem cell derived mice.

The Effect of Transcutaneous Electrical Acupoint Stimulation on Inflammatory Response in Patients Undergoing Limb Ischemia-Reperfusion

“…In addition, we found that IL-10 levels were not different between time points or groups. Guo et al found a similar result in a rat model of acute pancreatitis [ 39 ]. This suggests that TEAS may play a critical role in anti-inflammation through reducing the levels of proinflammatory cytokines in the plasma but may not have much influence on anti-inflammatory cytokines.…”

Contribution of GFP Expressing Dermal Papillae Cells to the Formation of Chimeric Embryos and their Survival in Uterine Environment

“…Earlier experiments in the field of cell reprogramming demonstrate that changes of the differentiation level in specialized epidermal cells can be achieved by nuclear transfer or under the influence of four specific genes regulating cell pluripotency [2,4,18]. There is evidence that induced pluripotent dermal fibroblast can be involved in the formation of teratomas in timusfree mouse [23] and defrosted GFP-MEFs derived iPSCs can participate in organ structures and the tissue building of chimera mouse [24]. Other research demonstrates substantial differences between adult stem-like cells and ESCs after the separation of mixed cell culture into GFP+/GFP-cell populations by flow cytometry and/or using mixed embryonic bodies to transfer under kidney capsules to create teratomes [22].…”

“…Previous basic research revealed that dermal macroenvironment is important in the maintenance of bulb cell population and hair follicle growth [21,22]. Based on findings that demonstrate the plasticity as well as regenerative and inductive properties of hair follicle dermal cells [23,24], we have postulated that their epigenetic can be changed under the influence of an embryonic microenvironment when DPCs are injected into the cavity of an expanded blastocyst. They may multiply and contribute to organs and tissue of mosaic animal models after transferring microinjected embryos to foster mothers.…”

Contribution of rat embryonic stem cells to xenogeneic chimeras in blastocyst or 8‐cell embryo injection and aggregation

“…Although the aggregation method requires less equipment and is easier to operate, it has the disadvantage that the embryo development stages that can be used are limited. Both methods have been widely used to generate transgenic mice, and differences in the generation and germ‐line transmission efficiency have been reported for each method . It was reported that the germ‐line transmission efficiency of chimeric animals increased by using 8‐cell embryos.…”

“…Both methods have been widely used to generate transgenic mice, and differences in the generation and germ-line transmission efficiency have been reported for each method. 17 It was reported that the germ-line transmission efficiency of chimeric animals increased by using 8-cell embryos.…”