Hyun Soo Kim, Cheol June Choi, Jin Yong Lee, Ho Young Ji, Kyu Seek Kim, Thida Win, Jong Ho Lee, Hyuk jin Moon, Dong Hyun Lee, Hyo Gyeong Jang, Soo Dong Woo
Kyu Seek Kim, Jun Su Bae, Hyun Soo Kim, Cheol June Choi, Jin Yong Lee, Ho Young Ji, Jong Ho Lee, Hyuk jin Moon, Hyo gyeong Jang, Dong hyun Lee, Seul Ki Kim, Do Young Kim, Jin Woong Im, Soo Dong Woo
The baculovirus expression system is a very useful tool widely used for expression of foreign proteins. To use the baculovirus expression system, a recombinant baculovirus must be prepared. The development of the Bac to Bac system has reduced the time and effort required to produce recombinant baculovirus. But, it will take at least two weeks. Further, it takes more time to measure the activity of recombinant baculovirus. In order to overcome this problem, a virus inducible expression system is being studied recently. Although baculovirus is able to rapidly express foreign proteins, it still has a low expression level. Thus, in this study, we aimed to construct a novel baculovirus inducible expression vector that not only shortens the production time of protein but also can express at a high level. The novel baculovirus inducible expression vector has been evaluated using EGFP and is expected to be a very useful tool for production of various proteins.
Neural stem cells (NSCs) are self-renewing tripotent cell populations and have capacity of neuronal (neurons) and glial (astrocytes and oligodendrocytes) differentiation. Many researchers have reported that NSCs have therapeutic effects in neurological disease by transplantation. However, it is not easy to obtain NSCs in vitro. Recently, Yamanaka and colleagues showed that somatic cells could be reprogrammed into pluripotent state by enforcing reprogramming factors. Induced pluripotent stem (iPS) cells undergo unlimited self-renewal and have differentiation potential into various types of cells like embryonic stem cells. Direct differentiation into a specialized cell types from iPS cells hold considerable promise for regenerative medicine as well as basic research. Here, we induced differentiation of iPS cells into NSCs in vitro and in vivo, which were compared with embryonic stem (ES) cell-derived NSCs and brain derived NSCs. NSCs from ES and iPS cells were morphologically indistinguishable from brain derived NSCs and stained positive for NSCs markers Nestin and Sox2. ES cells derived NSCs were transcriptionally distinguishable from brain derived NSCs. However, global gene expression pattern were similar but distinct between iPS derived NSCs and brain derived NSCs. Moreover, iPS derived NSCs were spontaneously aggregated upon passaging, formed ES cell like colonies, and finally reactivated Oct4-GFP. The spontaneously reverted GFP-positive cells (iPS-NSC-iPS) expressed similar levels of pluripotency markers (Oct4,Nanog) to ES and iPS cells, and could form germ line chimera. One possible explanation for this phenomenon is that spontaneously re-reprogramming was associated with transgene re-activation when iPS cells were differentiated into NSCs. However, NSCs from dox-inducible iPScells could not be reprogrammed into pluripotent state without doxycycline. Taken together, iPS derived NSCs were morphologically and similar to brain derived NSCs, but differ in gene expression pattern and maintenance. * This work was supported by the Next Generation Bio-Green21 Program funded by the Rural Development Administration (Grant PJ008009).
EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to 1000 ㎍/㎖ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of 50 ㎍/㎖ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of 50 ㎍/㎖. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.