Chanseok Lee

Isolation and initial culture of porcine inner cell masses derived from in vitro -produced blastocysts

The present study was conducted to isolate and culture inner cell mass (ICM) primarily derived from in vitro-produced blastocysts and to develop the culture conditions for the ICM cells. In Experiment 1, immunosurgically isolated ICMs of blastocysts derived from in vitro fertilization (IVF), somatic cell nuclear transfer (SCNT) or parthenogenetic activation (PA) were seeded onto STO cells. Primary colonies from each isolated ICM were formed with a ratio of 28.9, 30.0 and 4.9%, respectively. In Experiment 2, blastocysts collected from IVF were directly seeded onto a feeder layer with or without zona pellucida (ZP), or were subjected to ICM isolation by immunosurgery. Primary colonies were formed in 36.8% of isolated ICMs and 19.4% in intact blastocysts without ZP. In Experiment 3, ICMs from IVF blastocysts were seeded onto STO cells, mouse embryonic fibroblast (MEF) or porcine uterine epithelial cells (PUEC). On STO and MEF cells, 34.5 and 22.2% of primary colonies were formed, respectively. However, no primary colony was formed on the PUEC or in feeder-free condition. In Experiment 4, ICMs from IVF blastocysts were cultured in DMEM + Hams F10 (D/H medium), DMEM + NCSU-23 (D/N medium) or DMEM alone. When D/H medium or D/N medium was used, 21.7 or 44.4% of primary colony were formed, respectively, while no primary colony was formed in DMEM alone. These cells showed alkaline phosphatase activity and could be maintained for up to five passages. In suspension culture, cells formed embryoid bodies. These results demonstrate that porcine ICM could be isolated and cultured primarily from in vitro-produced blastocysts with a suitable culture system.

Robust microzip fastener: repeatable interlocking using polymeric rectangular parallelepiped arrays.

We report a highly repeatable and robust microzip fastener based on the van der Waals force-assisted interlocking between rectangular parallelepiped arrays. To investigate zipperlike interlocking behaviors, various line arrays were fabricated with three different spacing ratios (1, 3, and 5 of 800 nm in width) and width of parallelepipeds (400 nm, 800 nm, and 5 μm with the spacing ratio of 1). In addition, the different rigidity of line arrays was inspected for a repeatable microzip fastener. The normal and shear locking forces were measured with variation of the material rigidity as well as geometry of the array, in good agreement with a proposed theory based on the contact area and force balance. The maximum adhesion forces as high as ∼8.5 N cm(-2) in the normal direction and ∼29.6 N cm(-2) in the shear direction were obtained with high stability up to 1000 cycles. High stability of our fastening system was confirmed for preventing critical failures such as buckling and fracture in practical applications.

Repeated shape recovery of clustered nanopillars by mechanical pulling

High-aspect-ratio (HAR) nanopillars are of interest for wetting, adhesion, and energy harvesting due to their superior surface properties, including large surface area and high compliance. However, their intrinsically low mechanical stability has been a major obstacle for practical applications that require repeated use and in wet and humid environments. Herein, we show a method that can recover the clustered or deformed HAR nanopillars to their original shapes by taking advantage of the mechanical compliance of the nanopillars toward pulling during a demolding process. The pillars can be repeatedly clustered and recovered many times. Our method is simple yet powerful to recover the clustered nanopillars over a large area (7 × 10 cm2). By taking advantage of the different optical properties of the clustered pillars vs. the straight ones, we demonstrate display and erasing of patterns and tunable wettability by stamping the nanopillars to induce clustering, followed by shape recovery via demolding of the pillars.

Publisher Correction: Polymorphic design of DNA origami structures through mechanical control of modular components

The originally published version of this Article contained an error in Figure 5. In panel f, the right y-axis ‘Strain energy (kbT)’ was labelled ‘Probability’ and the left y-axis ‘Probability’ was labelled ‘Strain energy (kbT)’. This error has now been corrected in both the PDF and HTML versions of the Article.

Polymorphic design of DNA origami structures through mechanical control of modular components

Scaffolded DNA origami enables the bottom-up fabrication of diverse DNA nanostructures by designing hundreds of staple strands, comprised of complementary sequences to the specific binding locations of a scaffold strand. Despite its exceptionally high design flexibility, poor reusability of staples has been one of the major hurdles to fabricate assorted DNA constructs in an effective way. Here we provide a rational module-based design approach to create distinct bent shapes with controllable geometries and flexibilities from a single, reference set of staples. By revising the staple connectivity within the desired module, we can control the location, stiffness, and included angle of hinges precisely, enabling the construction of dozens of single- or multiple-hinge structures with the replacement of staple strands up to 12.8% only. Our design approach, combined with computational shape prediction and analysis, can provide a versatile and cost-effective procedure in the design of DNA origami shapes with stiffness-tunable units.The use of staple strands paired with scaffold strands allows the creation of a diverse array of DNA origami nanostructures. Here the authors rationally design a set of staples with variable hinges allowing controllable geometry and flexibility of the final structure.