Conversion of mouse fibroblasts to sphere cells induced by AlbuMAXI-containing medium

Wang

et al. 2018

IJMS

Large numbers of lipids exist in the porcine oocytes and early embryos and have the positive effects on their development, suggesting that the lipids may play an important role in pluripotency establishment and maintenance in pigs. However, the effects of lipids and their metabolites, such as fatty acids on reprogramming and the pluripotency gene expression of porcine-induced pluripotent stem cells (iPSCs), are unclear. Here, we generated the porcine iPSCs that resemble the mouse embryonic stem cells (ESCs) under lipid and fatty-acid-enriched cultural conditions (supplement of AlbuMAX). These porcine iPSCs show positive for the ESCs pluripotency markers and have the differentiation abilities to all three germ layers, and importantly, have the capability of aggregation into the inner cell mass (ICM) of porcine blastocysts. We further confirmed that lipid and fatty acid enriched condition can promote the cell proliferation and improve reprogramming efficiency by elevating cAMP levels. Interestingly, this lipids supplement promotes mesenchymal–epithelial transition (MET) through the cAMP/PKA/CREB signal pathway and upregulates the E-cadherin expression during porcine somatic cell reprogramming. The lipids supplement also makes a contribution to lipid droplets accumulation in the porcine iPSCs that resemble porcine preimplantation embryos. These findings may facilitate understanding of the lipid metabolism in porcine iPSCs and lay the foundation of bona fide porcine embryonic stem cell derivation.

Section: Introductionmentioning

confidence: 99%

Lipid Supplement in the Cultural Condition Facilitates the Porcine iPSC Derivation through cAMP/PKA/CREB Signal Pathway

Wang

et al. 2018

IJMS

“…Mouse fibroblast cells were transdifferentiated into an intermediate state characterized by spherical cell aggregates, somewhere between mouse embryonic stem cells and mouse fibroblasts, using specialized medium containing lipid-rich albumin. Such spheres were able to differentiate into ectodermal germ layers following retinoic acid treatment[20]. microRNAs, such as miR9* and miR-124, participated in the transdifferentiation of fibroblasts into neuronal cells in the presence of other neurogenic transcription factors[2122].…”

Section: Introductionmentioning

confidence: 99%

Mash1 efficiently reprograms rat astrocytes into neurons

Ding

et al. 2014

Neural Regen Res

To date, it remains poorly understood whether astrocytes can be easily reprogrammed into neurons. Mash1 and Brn2 have been previously shown to cooperate to reprogram fibroblasts into neurons. In this study, we examined astrocytes from 2-month-old Sprague-Dawley rats, and found that Brn2 was expressed, but Mash1 was not detectable. Thus, we hypothesized that Mash1 alone could be used to reprogram astrocytes into neurons. We transfected a recombinant MSCV-MASH1 plasmid into astrocytes for 72 hours, and saw that all cells expressed Mash1. One week later, we observed the changes in morphology of astrocytes, which showed typical neuronal characteristics. Moreover, β-tubulin expression levels were significantly higher in astrocytes expressing Mash1 than in control cells. These results indicate that Mash1 alone can reprogram astrocytes into neurons.

“…During the same stage of reprogramming, NCs showed increased sterol biosynthesis (Fig ). In a previous study, murine embryonic fibroblasts could be transformed into sphere cells by suspension culture in lipid‐rich medium (Rajanahalli et al , ). Therefore, we theorized that the period prior to the MET process may represent a critical period for the removal of cell identity and that changing patterns of gene expression may provide insights into trans‐differentiation processes of different somatic cells.…”

Section: Discussionmentioning

confidence: 99%

Reprogramming‐derived gene cocktail increases cardiomyocyte proliferation for heart regeneration

et al. 2016

Although remnant cardiomyocytes (CMs) possess a certain degree of proliferative ability, efficiency is too low for cardiac regeneration after injury. In this study, we identified a distinct stage within the initiation phase of CM reprogramming before the MET process, and microarray analysis revealed the strong up‐regulation of several mitosis‐related genes at this stage of reprogramming. Several candidate genes were selected and tested for their ability to induce CM proliferation. Delivering a cocktail of three genes, FoxM1, Id1, and Jnk3‐shRNA (FIJs), induced CMs to re‐enter the cell cycle and complete mitosis and cytokinesis in vitro. More importantly, this gene cocktail increased CM proliferation in vivo and significantly improved cardiac function and reduced fibrosis after myocardial infarction. Collectively, our findings present a cocktail FIJs that may be useful in cardiac regeneration and also provide a practical strategy for probing reprogramming assays for regeneration of other tissues.