Generation of Chicken Contractile Skeletal Muscle Structure Using Decellularized Plant Scaffolds.

Cultured meat is a meat analogue produced by in vitro cell culture, which can replace the conventional animal production system. Tissue engineering using myogenic cells and biomaterials is a core technology for cultured meat production. In this study, we provide an efficient and economical method to produce skeletal muscle tissue-like structures by culturing chicken myoblasts in a fetal bovine serum (FBS)-free medium and plant-derived scaffolds. An FBS-free medium supplemented with 10% horse serum (HS) and 5% chick embryo extract (CEE) was suitable for the proliferation and differentiation of chicken myoblasts. Decellularized celery scaffolds (Decelery), manufactured using 1% sodium dodecyl sulfate (SDS), were nontoxic to cells and supported myoblast proliferation and differentiation. Decelery could support the 3D culture of chicken myoblasts, which could adhere and coagulate to the surface of the Decelery and form MYH1E+ and F-actin+ myotubes. After 2 weeks of culture on Decelery, fully grown myoblasts completely covered the surface of the scaffolds and formed fiber-like myotube structures. They further differentiated to form spontaneously contracting myofiber-like myotubes on the scaffold surface, indicating that the Decelery scaffold system could support the formation of a functional mature myofiber structure. In addition, as the spontaneously contracting myofibers did not detach from the surface of the Decelery, the Decelery system is a suitable biomaterial for the long-term culture and maintenance of the myofiber structures.

Altered Mitochondrial Function and Accelerated Aging Phenotype in Neural Stem Cells Derived from Dnm1l Knockout Embryonic Stem Cells.

Mitochondria are crucial for cellular energy metabolism and are involved in signaling, aging, and cell death. They undergo dynamic changes through fusion and fission to adapt to different cellular states. In this study, we investigated the effect of knocking out the dynamin 1-like protein (Dnm1l) gene, a key regulator of mitochondrial fission, in neural stem cells (NSCs) differentiated from Dnm1l knockout embryonic stem cells (Dnm1l-/- ESCs). Dnm1l-/- ESC-derived NSCs (Dnm1l-/- NSCs) exhibited similar morphology and NSC marker expression (Sox2, Nestin, and Pax6) to brain-derived NSCs, but lower Nestin and Pax6 expression than both wild-type ESC-derived NSCs (WT-NSCs) and brain-derived NSCs. In addition, compared with WT-NSCs, Dnm1l-/- NSCs exhibited distinct mitochondrial morphology and function, contained more elongated mitochondria, showed reduced mitochondrial respiratory capacity, and showed a metabolic shift toward glycolysis for ATP production. Notably, Dnm1l-/- NSCs exhibited impaired self-renewal ability and accelerated cellular aging during prolonged culture, resulting in decreased proliferation and cell death. Furthermore, Dnm1l-/- NSCs showed elevated levels of inflammation and cell stress markers, suggesting a connection between Dnm1l deficiency and premature aging in NSCs. Therefore, the compromised self-renewal ability and accelerated cellular aging of Dnm1l-/- NSCs may be attributed to mitochondrial fission defects.

Embryonic Stem Cells Lacking DNA Methyltransferases Differentiate into Neural Stem Cells that Are Defective in Self-Renewal.

Background and Objectives: DNA methyltransferases (Dnmts) play an important role in regulating DNA methylation during early developmental processes and cellular differentiation. In this study, we aimed to investigate the role of Dnmts in neural differentiation of embryonic stem cells (ESCs) and in maintenance of the resulting neural stem cells (NSCs). Methods and Results: We used three types of Dnmt knockout (KO) ESCs, including Dnmt1 KO, Dnmt3a/3b double KO (Dnmt3 DKO), and Dnmt1/3a/3b triple KO (Dnmt TKO), to investigate the role of Dnmts in neural differentiation of ESCs. All three types of Dnmt KO ESCs could form neural rosette and differentiate into NSCs in vitro. Interestingly, however, after passage three, Dnmt KO ESC-derived NSCs could not maintain their self-renewal and differentiated into neurons and glial cells. Conclusions: Taken together, the data suggested that, although deficiency of Dnmts had no effect on the differentiation of ESCs into NSCs, the latter had defective maintenance, thereby indicating that Dnmts are crucial for self-renewal of NSCs.

Developmental Potency and Metabolic Traits of Extended Pluripotency Are Faithfully Transferred to Somatic Cells via Cell Fusion-Induced Reprogramming.

As a novel cell type from eight-cell-stage embryos, extended pluripotent stem cells (EPSCs) are known for diverse differentiation potency in both extraembryonic and embryonic lineages, suggesting new possibilities as a developmental research model. Although various features of EPSCs have been defined, their ability to directly transfer extended pluripotency to differentiated somatic cells by cell fusion remains to be elucidated. Here, we derived EPSCs from eight-cell mouse embryos and confirmed their extended pluripotency at the molecular level and extraembryonic differentiation ability. Then, they were fused with OG2+/- ROSA+/- neural stem cells (NSCs) by the polyethylene-glycol (PEG)-mediated method and further analyzed. The resulting fused hybrid cells exhibited pluripotential markers with upregulated EPSC-specific gene expression. Furthermore, the hybrid cells contributed to the extraembryonic and embryonic lineages in vivo and in vitro. RNA sequencing analysis confirmed that the hybrid cells showed distinct global expression patterns resembling EPSCs without parental expression of NSC markers, indicating the complete acquisition of extended pluripotency and the erasure of the somatic memory of NSCs. Furthermore, ultrastructural observation and metabolic analysis confirmed that the hybrid cells rearranged the mitochondrial morphology and bivalent metabolic profile to those of EPSCs. In conclusion, the extended pluripotency of EPSCs could be transferred to somatic cells through fusion-induced reprogramming.

Meiosis-specific cohesin complexes display essential and distinct roles in mitotic embryonic stem cell chromosomes.

BACKGROUND: Cohesin is a chromosome-associated SMC-kleisin complex that mediates sister chromatid cohesion, recombination, and most chromosomal processes during mitosis and meiosis. However, it remains unclear whether meiosis-specific cohesin complexes are functionally active in mitotic chromosomes. RESULTS: Through high-resolution 3D-structured illumination microscopy (3D-SIM) and functional analyses, we report multiple biological processes associated with the meiosis-specific cohesin components, α-kleisin REC8 and STAG3, and the distinct loss of function of meiotic cohesin during the cell cycle of embryonic stem cells (ESCs). First, we show that STAG3 is required for the efficient localization of REC8 to the nucleus by interacting with REC8. REC8-STAG3-containing cohesin regulates topological properties of chromosomes and maintains sister chromatid cohesion. Second, REC8-cohesin has additional sister chromatid cohesion roles in concert with mitotic RAD21-cohesin on ESC chromosomes. SIM imaging of REC8 and RAD21 co-staining revealed that the two types of α-kleisin subunits exhibited distinct loading patterns along ESC chromosomes. Third, knockdown of REC8 or RAD21-cohesin not only leads to higher rates of premature sister chromatid separation and delayed replication fork progression, which can cause proliferation and developmental defects, but also enhances chromosome compaction by hyperloading of retinoblastoma protein-condensin complexes from the prophase onward. CONCLUSIONS: Our findings indicate that the delicate balance between mitotic and meiotic cohesins may regulate ESC-specific chromosomal organization and the mitotic program.

Germ Cell Derivation from Pluripotent Stem Cells for Understanding In Vitro Gametogenesis.

Assisted reproductive technologies (ARTs) have developed considerably in recent years; however, they cannot rectify germ cell aplasia, such as non-obstructive azoospermia (NOA) and oocyte maturation failure syndrome. In vitro gametogenesis is a promising technology to overcome infertility, particularly germ cell aplasia. Early germ cells, such as primordial germ cells, can be relatively easily derived from pluripotent stem cells (PSCs); however, further progression to post-meiotic germ cells usually requires a gonadal niche and signals from gonadal somatic cells. Here, we review the recent advances in in vitro male and female germ cell derivation from PSCs and discuss how this technique is used to understand the biological mechanism of gamete development and gain insight into its application in infertility.

Current Issues and Technical Advances in Cultured Meat Production: A Review.

As the global population grows, we need a stable protein supply to meet the demands. Although plant-derived protein sources are widely available, animal meat maintains its popularity as a high-quality and savory protein source. Recently, cultured meat, also known as in vitro meat, has been suggested as a meat analog produced through in vitro cell culture technology. Cultured meat has several advantages over conventional meat, such as environmental protection, disease prevention, and animal welfare. However, cultured meat manufacturing is an emerging technology; thus, its further and dynamic development would be pivotal. Commercialization of cultured meat to the public will take a long time but cultured meat undoubtedly will come to our table someday. Here, we discuss the social and economic aspects of cultured meat production as well as the recent technical advances in cultured meat technology.

Predictive factors of long-term follow-up in treatment of Korean alcoholics with naltrexone or acamprosate.

Medication is integral in treating alcohol use disorder. However, because of the small to medium effect size, much effort is being exerted to identify predicting factors for effective pharmacological treatment in alcohol abuse disorder. Rather than focusing on abstinence days, alcohol craving, or frequency of drinking, which has been the focus of previous studies. Recently, there have been several studies which focused on follow-up length as an indicator of pharmacological treatment effe ctiveness. The purpose of this study was to investigate the predicting factors of long-term follow-up in treating Korean alcoholics with naltrexone or acamprosate. A retrospective study was conducted. Medical records of all patients diagnosed from November 2008 to May 2017 with alcohol abuse or alcohol dependence at psychiatric clinics at PNUYH were reviewed. We examined total days of which naltrexone or acamprosate were prescribed, and investigated if there were predicting factors maintaining follow-up at least 180 days or more. With these data, logistic regression analysis was conducted. In naltrexone long group compared to naltrexone short group, factors of having medical comorbidities [odds ratio (OR) = 5.477, P = 0.012] showed higher OR. In acamprosate long group, factors of age (OR = 1.083, P = 0.030), and use of more than four psychotropic medications (OR = 7.107, P = 0.030), showed higher OR. In both medications, predicting factors were different from the other. Further study to investigate the reasons would provide us with a new insight.

Seismic detection of strong ground motions by MW5.6 North Korean nuclear explosion.

The North Korean nuclear explosion test site in Punggye-ri is located in a seismically quiescent region on a stable Precambrian basement. The 3 September 2017 MW5.6 North Korean underground nuclear explosion (UNE) test produced unprecedented strong ground motions. The peak ground accelerations might reach tens to hundreds m/s2 on the surface of the UNE test site, decaying exponentially with distance. Ten shallow events with magnitudes greater than or equal to ML2.5 and source depths less than 3 km followed the 2017 UNE for 5 months in an area with a radius of 15 km from the UNE where strong ground shaking was experienced. The largest event with MW3.7 occurred 20 days after the 2017 UNE test at shallow depths less than 3 km. Its moment tensor solution indicates a combined source behavior with comparable strengths of double-couple and compensated linear vector dipole (CLVD) components, suggesting an unusual event different from typical natural earthquakes in the Korean Peninsula. The clustered shallow seismic events appeared to have occurred in damaged media that were effectively perturbed by the strong ground motions of the UNE.

Time-advanced occurrence of moderate-size earthquakes in a stable intraplate region after a megathrust earthquake and their seismic properties.

The distance-dependent coseismic and postseismic displacements produced by the 2011 MW9.0 Tohoku-Oki megathrust earthquake caused medium weakening and stress perturbation in the crust around the Korean Peninsula, increasing the seismicity with successive ML5-level earthquakes at the outskirts of high seismicity regions. The average ML5-level occurrence rate prior to the megathrust earthquake was 0.15 yr-1 (0.05-0.35 yr-1 at a 95% confidence level), and the rate has increased to 0.71 yr-1 (0.23-1.67 yr-1 at a 95% confidence level) since the megathrust earthquake. The 2016 ML5-level midcrustal earthquakes additionally changed the stress field in adjacent regions, inducing the 15 November 2017 ML5.4 earthquake. The successive 2016 and 2017 moderate-size earthquakes built complex stress fields in the southeastern Korean Peninsula, increasing the seismic hazard risks in the regions of long-term stress accumulation. The increased seismic risks may continue until the medium properties and stress field are recovered.

Prediction of ground motion and dynamic stress change in Baekdusan (Changbaishan) volcano caused by a North Korean nuclear explosion.

Strong ground motions induce large dynamic stress changes that may disturb the magma chamber of a volcano, thus accelerating the volcanic activity. An underground nuclear explosion test near an active volcano constitutes a direct treat to the volcano. This study examined the dynamic stress changes of the magma chamber of Baekdusan (Changbaishan) that can be induced by hypothetical North Korean nuclear explosions. Seismic waveforms for hypothetical underground nuclear explosions at North Korean test site were calculated by using an empirical Green's function approach based on a source-spectral model of a nuclear explosion; such a technique is efficient for regions containing poorly constrained velocity structures. The peak ground motions around the volcano were estimated from empirical strong-motion attenuation curves. A hypothetical M7.0 North Korean underground nuclear explosion may produce peak ground accelerations of 0.1684 m/s(2) in the horizontal direction and 0.0917 m/s(2) in the vertical direction around the volcano, inducing peak dynamic stress change of 67 kPa on the volcano surface and ~120 kPa in the spherical magma chamber. North Korean underground nuclear explosions with magnitudes of 5.0-7.6 may induce overpressure in the magma chamber of several tens to hundreds of kilopascals.