Since rice is the main food in Korea, there are no regulations on corn milling yet. Corn is known as one of the world's top three food crops along with wheat and rice, and it is known that 3.5 billion people worldwide use corn for food. In addition, corn mills are not developed or sold in Korea, but the use of corn mills is increasing significantly in many countries in Southeast Asia. In the Philippines, as Korea's rice mill import increases, Korea's KAMICO (Korea Agricultural Machinery Industry Cooperative) and domestic company A agreed to develop a corn mill jointly with PHilMech, an organization affiliated with the Philippine Ministry of Agriculture. However, research on corn milling was very insignificant, so the development was carried out based on the technology of Korea's rice mill. Rice milling is performed by peeling off the skin of rice and producing brown or white rice, so it is carried out by removing the skin and cutting the skin. On the other hand, in the corn mill, the skin of the corn is peeled, pulverized and selected to produce main products suitable for edible use. Therefore, in order to develop a corn mill, processes such as peeling, transfer, grinding, sorting, and by-product separation are required, and suitable parts must be developed. In addition, the performance must be gradually improved through experiments in which corn is repeatedly milled. The Philippines produces 7.98 million tons/year of corn, which is about 100 times that of Korea, and is mostly consumed as a staple food. This is about 10% of the total crop production in the Philippines. In addition, the main cultivation complexes of corn are the mountainous regions of Tarlac or Pangasinan, and the produced corn is 72.4% of the so-called yellow corn called Arabel and Sarangani, and the remaining 27.6% are known as white corn. In this study, it was intended to produce grains of 2.5 mm or less suitable for food for yellow corn and to develop a corn mill for 200 kg per hour. Detailed conditions for development are stipulated as more than 55% of the main product recovery rate, more than 31% of the by-product recovery rate, less than 5% of the raw material loss rate, and more than 80% of the embryo dislocation rate. In this study, to achieve this, the overall process of the corn mill was developed, and the optimal conditions for the corn mill were obtained through the development of parts and empirical tests to improve performance. In addition, it was intended to achieve the development goal by evaluating and analyzing the performance of each part so that it did not conflict.
원형질체 융합에 의한 화합성 및 불화합성 종간 체세포잡종을 얻었다. 화합성 종간인 Pleurotus ostreatus 와 P. florida 의 융합체는 이질핵체 (heterokaryon) 를 형성하였고, 불화합성 종간인 P. cornucopiae + P. florida , P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, 그리고 P. ostreatus + Flammulina velutipes 는 합핵체(synkaryon) 를 형성하였다. 이질이핵체는 동일한 양상의 자실체를 형성하는데 비해 합핵체는 유사분열상의 꺽쇠연결체 형성, 한쪽 친과 유사한 자실체 형성, 비정상적 유전형질 분리 및 유전자재조합 현상을 나타내었다. 화합성 및 불화합성 계통간 융합체의 RAPD 분석결과 화합성 종간 융합체는 동일한 DNA 패턴을 나타내었고, 불화합성 종간 융합체는 한쪽 친과 유사한 DNA 양상이면서 비양친 DNA 밴드도 형성하였다. 합핵체의 패턴은 microgenome insertion type 과 macrogenome insertion type 으로 구분되었다. 합핵체의 자실체 발생은 융합 모균주 양친의 자가임성에 의존하는데 이는 느타리의 동형핵체 자가임성과 유사한 양상이었고, 교배형 전환과 관련이 있는 것으로 사료된다. 여기서는 이러한 관점에서 논할 것이다.
Highly homogeneous and functional stem cell-derived hepatocyte-like cells (HLCs) are considered a promising option in the treatment of liver disease and the development of effective in vitro toxicity screening tool. However, the purity of cells and expression and/or activity of drug metabolizing enzymes in stem cell-derived HLCs are usually too low to be useful for clinical or in vitro applications. Here, we describe a highly optimized differentiation protocol, which produces more than 90% albumin-positive HLCs with no purification process. In addition, we show that hepatic enzyme gene expressions and activities were significantly improved by generating three-dimensional (3D) spheroidal aggregate of HLCs. The 3D differentiation method increased expressions of nuclear receptors that regulate the proper expression of key hepatic enzymes. Furthermore, a significantly increased hepatic functions such as albumin and urea secretion were observed in 3D hepatic spheroids and HLCs in the spheroid exhibited morphological and ultrastructural features of normal hepatocytes. Importantly, we show that repeated exposures to xenobiotics facilitated the functional maturation of HLC, as confirmed by increased expression of genes for drug metabolizing enzymes and transcription factors. In conclusion, the 3D culture system with repeated exposures to xenobiotics may be a new strategy for enhancing hepatic maturation of stem cell-derived HLCs as a cell source for in vitro high-throughput hepatotoxicity models.
Hepatocyte-like cells (HLCs) derived from human pluripotent stem cells have received extensive attention in the development of drug screening and toxicity testing. However, it has been reported that stem cell-derived HLCs showed hepatic functions that were too limited to be of use in drug screening and toxicity testing, possibly due to the lack of sufficient intercellular communication under conventional two-dimensional (2D) culture conditions. Therefore, a 3D differentiation system may overcome the in vitro limitation of 2D culture to produce stem cell-derived hepatocytes with mature metabolic functions. In this study, the feasibility of using a silicone-based spherofilm, specifically designed to produce spherical cell clusters, to generate uniformly sized 3D hepatic spheroids from hESCs was investigated. Hepatic spheroids generated on the spherofilm showed more homogenous size and shape than those generated in conventional low-attachment suspension culture dishes. Results of immunohistochemical analysis showed that expression of the mature hepatic marker albumin (ALB) increased over time during the hepatic maturation process. Furthermore, the 3D culture system mimicked the in vivo 3D microenvironment. Laminin, which is an important component of hepatic ECM, was expressed in hepatic spheroids. The results of immunohistochemical analysis indicated that the 3D culture environment is capable of generating an in vivo-like microenvironment. In addition, quantitative PCR analysis showed that the mature hepatic marker ALB and cytochrome P450 (CYP) enzymes CYP3A4 and CYP3A7 were expressed at higher levels in 3D culture than in 2D culture. This indicates that the 3D culture system is suitable for hepatic maturation and that our size-controlled 3D culture conditions might accelerate hepatic function. These results suggest that 3D hepatic spheroids significantly enhance metabolic maturation of hepatocytes derived from hESCs