Residual-Solvent-Induced Morphological Transformation by Intense Pulsed Light on Spin-Coated and Inkjet-Printed ZnO NP Films for Quantum-Dot Light-Emitting Diodes.

It was demonstrated through a comparison between the spin-coated and inkjet-printed quantum-dot light-emitting diodes' (QLED) performance analysis outcomes that the annealing temperature of a zinc oxide nanoparticle (ZnO NP) electron transport layer (ETL) optimized for intense pulsed light (IPL) via a post-treatment differs depending on the film-formation method used. For a naturally dried ZnO NP ETL formulated without annealing, different film morphologies were observed according to the film-formation method of spin coating and inkjet printing, and the surface-roughness root mean square (RMS) value was increased in an IPL post-treatment due to unevaporated residual solvent. Based on this phenomenon, we classified and analyzed different film profiles according to the deposition method, the presence or absence of annealing, and the annealing temperature.

Investigation of the Chemical Structure of Ultra-Thin Polyimide Substrate for the Xenon Flash Lamp Lift-off Technology.

Lift-off is one of the last steps in the production of next-generation flexible electronics. It is important that this step is completed quickly to prevent damage to ultrathin manufactured electronics. This study investigated the chemical structure of polyimide most suitable for the Xe Flash lamp-Lift-Off process, a next-generation lift-off technology that will replace the current dominant laser lift-off process. Based on the characteristics of the peeled-off polyimide films, the Xe Flash lamp based lift-off mechanism was identified as photothermal decomposition. This occurs by thermal conduction via light-to-heat conversion. The synthesized polyimide films treated with the Xe Flash lamp-Lift-Off process exhibited various thermal, optical, dielectric, and surface characteristics depending on their chemical structures. The polyimide molecules with high concentrations of -CF3 functional groups and kinked chemical structures demonstrated the most promising peeling properties, optical transparencies, and dielectric constants. In particular, an ultra-thin polyimide substrate (6 µm) was successfully fabricated and showed potential for use in next-generation flexible electronics.

Effect of Time-Dependent Characteristics of ZnO Nanoparticles Electron Transport Layer Improved by Intense-Pulsed Light Post-Treatment on Hole-Electron Injection Balance of Quantum-Dot Light-Emitting Diodes.

We investigated the effect of intense-pulsed light (IPL) post-treatment on the time-dependent characteristics of ZnO nanoparticles (NPs) used as an electron transport layer (ETL) of quantum-dot light-emitting diodes (QLEDs). The time-dependent characteristics of the charge injection balance in QLEDs was observed by fabrication and analysis of single carrier devices (SCDs), and it was confirmed that the time-dependent characteristics of the ZnO NPs affect the device characteristics of QLEDs. Stabilization of the ZnO NPs film properties for improvement of the charge injection balance in QLEDs was achieved by controlling the current density characteristics via filling of the oxygen vacancies by IPL post-treatment.

Sequential Improvement from Cosolvents Ink Formulation to Vacuum Annealing for Ink-Jet Printed Quantum-Dot Light-Emitting Diodes.

Optimization of ink-jet printing conditions of quantum-dot (QD) ink by cosolvent process and improvement of quantum-dot light-emitting diodes (QLEDs) characteristics assisted by vacuum annealing were analyzed in this research. A cosolvent process of hexane and ortho-dichlorobenzene (oDCB) was optimized at the ratio of 1:2, and ink-jetting properties were analyzed using the Ohnesorge number based on the parameters of viscosity and surface tension. However, we found that these cosolvents systems cause an increase in the boiling point and a decrease in the vapor pressure, which influence the annealing characteristics of the QD emission layer (EML). Therefore, we investigated QLEDs' performance depending on the annealing condition for ink-jet printed QD EML prepared using cosolvents systems of hexane and oDCB. We enhanced the quality of QD EML and device performance of QLEDs by a vacuum annealing process, which was used to prevent exposure to moisture and oxygen and to promote effective evaporation of solvent in QD EML. As a result, the characteristics of QLEDs formed using ink-jet printed QD EML annealed under vacuum environment increased luminescence (L), current efficiency (CE), external quantum efficiency (EQE), and lifetime (LT50) by 30.51%, 33.7%, 21.70%, and 181.97%, respectively, compared to QLEDs annealed under air environment.

Xenon Flash Lamp Lift-Off Technology without Laser for Flexible Electronics.

This study experimentally investigated process mechanisms and characteristics of newly developed xenon flash lamp lift-off (XF-LO) technology, a novel thin film lift-off method using a light to heat conversion layer (LTHC) and a xenon flash lamp (XFL). XF-LO technology was used to lift-off polyimide (PI) films of 8.68-19.6 µm thickness. When XFL energy irradiated to the LTHC was 2.61 J/cm2, the PI film was completely released from the carrier substrate. However, as the energy intensity of the XFL increased, it became increasingly difficult to completely release the PI film from the carrier substrate. Using thermal gravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FTIR) and transmittance analysis, the process mechanism of XF-LO technology was investigated. Thermal durability of the PI film was found to deteriorate with increasing XFL energy intensity, resulting in structural deformation and increased roughness of the PI film surface. The optimum energy intensity of 2.61 J/cm2 or less was found to be effective for performing XF-LO technology. This study provides an attractive method for manufacturing flexible electronic boards outside the framework of existing laser lift-off (LLO) technology.

Optical and Electrical Analysis of Annealing Temperature of High-Molecular Weight Hole Transport Layer for Quantum-dot Light-emitting Diodes.

In this study, we introduce optimization of the annealing conditions for improvement of hardness and hole transporting properties of high-molecular weight poly [9, 9-dioctylfluorene-co-N-(4-(3-methylpropyl)) diphenylamine] (TFB) film used as a Hole Transport Layer (HTL) of Quantum-dot Light-emitting Diodes (QLEDs). As annealing temperatures were increased from 120 °C to 150 °C or more, no dissolving or intermixing phenomena at the interface between HTL and Quantum-Dot Emission Layer (QDs EML) was observed. However, when the annealing temperatures was increased from 150 °C to 210 °C, the intensity of the absorbance peaks as determined by Fourier Transform Infrared (FT-IR) measurement was found to relatively decrease, and hole transporting properties were found to decrease in the measurement of current density - voltage (CD - V) and capacitance - voltage (C - V) characteristics of Hole Only Devices (HODs) due to thermal damage. At the annealing temperature of 150 °C, the QLEDs device was optimized with TFB films having good hardness and best hole transporting properties for solution processed QLEDs.

Contactless, nondestructive determination of dopant profiles of localized boron-diffused regions in silicon wafers at room temperature.

We develop a photoluminescence-based technique to determine dopant profiles of localized boron-diffused regions in silicon wafers and solar cell precursors employing two excitation wavelengths. The technique utilizes a strong dependence of room-temperature photoluminescence spectra on dopant profiles of diffused layers, courtesy of bandgap narrowing effects in heavily-doped silicon, and different penetration depths of the two excitation wavelengths in silicon. It is fast, contactless, and nondestructive. The measurements are performed at room temperature with micron-scale spatial resolution. We apply the technique to reconstruct dopant profiles of a large-area (1 cm × 1 cm) boron-diffused sample and heavily-doped regions (30 µm in diameter) of passivated-emitter rear localized-diffused solar cell precursors. The reconstructed profiles are confirmed with the well-established electrochemical capacitance voltage technique. The developed technique could be useful for determining boron dopant profiles in small doped features employed in both photovoltaic and microelectronic applications.

Investigation of Intrinsic Electrical Characteristics and Contact Effects in p-Type Tin Monoxide Thin-Film Transistors Using Gated-Four-Probe Measurements.

We investigate the intrinsic electrical characteristics and source/drain parasitic resistance in p-type SnO TFTs fabricated using Ni electrodes based on the gated-four-probe method. Because of the relatively high work function and inexpensive price, Ni has been most frequently used as the source/drain electrode materials in p-type SnO TFTs. However, our experimental data shows that the width normalized parasitic resistances of SnO TFT with Ni electrodes are around one to three orders of magnitude higher than those in the representative n-type oxide TFT, amorphous indium- gallium-zinc oxide TFT, and are comparable with those in amorphous silicon TFTs with Mo electrodes. This result implies that the electrical performance of the short channel SnO TFT can be dominated by the source/drain parasitic resistances. The intrinsic field-effect mobility extracted without being influenced by source/drain parasitic resistance was ~2.0 cm2/Vs, which is around twice the extrinsic field-effect mobility obtained from the conventional transconductance method. The large contact resistance is believed to be mainly caused from the heterogeneous electronic energy-level mismatch between the SnO and Ni electrodes.

Effect of Population Reduction on mtDNA Diversity and Demographic History of Korean Cattle Populations.

The population sizes of three Korean indigenous cattle populations have been drastically reduced over the past decades. In this study, we examined the extent to which reduction in populations influenced genetic diversity, population structure and demographic history using complete mitochondrial DNA (mtDNA) control region sequences. The complete mtDNA control region was sequenced in 56 individuals from Korean Black (KB), Jeju Black (JEB) and Korean Brindle (BRI) cattle populations. We included 27 mtDNA sequences of Korean Brown (BRO) from the GenBank database. Haplotype diversity estimate for the total population was high (0.870) while nucleotide diversity was low (0.004). The KB showed considerably low nucleotide (π = 0.001) and haplotype (h = 0.368) diversities. Analysis of molecular variance revealed a low level of genetic differentiation but this was highly significant (p<0.001) among the cattle populations. Of the total genetic diversity, 7.6% was attributable to among cattle populations diversity and the rest (92.4%) to differences within populations. The mismatch distribution analysis and neutrality tests revealed that KB population was in genetic equilibrium or decline. Indeed, unless an appropriate breeding management practice is developed, inbreeding and genetic drift will further impoverish genetic diversity of these cattle populations. Rational breed development and conservation strategy is needed to safeguard these cattle population.

Generation of functional cardiomyocytes from mouse induced pluripotent stem cells.

BACKGROUND: Induced pluripotent stem (iPS) cells allow derivation of autologous differentiated cells for cell therapy. The purpose of this study was to compare the cardiac differentiation potential of mouse iPS cells with embryonic stem (ES) cells and demonstrate that they could produce functional cardiomyocytes. METHODS: iPS cells were prepared from mouse embryonic fibroblasts by lentiviral mediated expression of four transcription factors (Oct4/Sox2/Klf4/C-myc). To induce cardiac cell differentiation, iPS-S-6 or D3-ES cells were induced to form embryoid bodies (EBs) using a two-medium culture protocol, then plated onto gelatin-coated plates and maintained in DMEM. RESULTS: Following classification of the generation periods of contracting EBs into early (d8-d11), middle (d12-d15) and late (d16-20), iPS cells in the early period exhibited characteristics similar to ES cells. In iPS cells from the middle period group, the ratio of contracting EBs was significantly increased compared to ES cells, and the difference persisted in cells from the late period group (p<0.05). The percentage of contracting EBs formed from iPS and ES cells were 44.8% and 33.3%, respectively. In addition, iPS cell-derived cardiomyocytes exhibited mRNA expression of cardiac mesoderm markers such as GATA4 and NKX2.5, and cardiomyocyte markers such as α1s, α1c, α-MHC, ß-MHC, Cx40, TnI, TnT, ANF and Hey2. Single cardiomyocytes exhibited typical cross-striated myofibrillar organization, and electrophysiological studies revealed functional cardiac-specific voltage-gated Na(+), Ca(2+) and K(+) channels. CONCLUSIONS: These results demonstrate that functional cardiomyocytes can be generated from iPS cells, and suggest that these cells may be useful for the treatment of cardiovascular disease.

3,4-Dihydroxyflavone acts as an antioxidant and antiapoptotic agent to support bovine embryo development in vitro.

The effects of two antioxidants, superoxide dismutase (SOD) and the flavonoid 3,4-dihydroxyflavone (DHF), on bovine embryo development in vitro were examined. Blastocyst development, total cell and inner cell mass (ICM) numbers, intracellular levels of reactive oxygen species (ROS), apoptotic indices and gene expression levels were examined before and after treatment of day 2 bovine embryos (≥2-4 cells) with various concentrations of 3,4-DHF or SOD for 6 days. Statistical analysis was performed using analysis of variance, with significance defined at the P<0.05 level. SOD had no significant effect on bovine embryo development at any tested concentration (control, 32.8%; 300 U/ml, 33.9%; 600 U/ml, 24.2%). In contrast, 10 µM 3,4-DHF promoted higher blastocyst development (39.3%) than any other concentration (control, 26.7%; 1 µM, 30.3%; 50 µM, 29.5%; 100 µM, 20.5%). Compared with 300 U/ml SOD, 10 µM 3,4-DHF resulted in significantly higher blastocyst development (44.2%) (control, 31.5%; SOD 300 U/ml, 33.6%). Treatment with 3,4-DHF increased the ICM cell number and reduced intracellular ROS production and apoptotic cell numbers. When O(2) tension was decreased from 20% (high tension) to 5% (low tension), embryo development rates were doubled regardless of 3,4-DHF treatment. Under high O(2) tension, 10 µM 3,4-DHF treatment may render bovine embryo development similar to a low O(2) tension environment. The best blastocyst development was obtained under low O(2) tension plus 10 µM 3,4-DHF treatment. The relative expression levels of antioxidant (MnSOD), antiapoptotic (Survivin, Bax inhibitor) and growth-related genes (IFN-τ, Glut-5) were significantly increased after 3,4-DHF treatment, while the expression levels of oxidant (Sox) and apoptotic genes (Caspase-3 and Bax) were reduced. These results suggest that 3,4-DHF may promote the in vitro development of bovine embryos through its antioxidant and antiapoptotic effects.

Differences between cellular and molecular profiles of induced pluripotent stem cells generated from mouse embryonic fibroblasts.

Induced pluripotent stem (iPS) cells are a new alternative for the development of patient-specific stem cells, and the aim of this study was to determine whether differences exist between the cellular and molecular profiles of iPS cells, generated using lentiviral vectors, compared to ES cells. The lentiviral infection efficiency differed according to the method of cell culture (adherent cells: 0.085%; suspended cells: 0.785%). Six iPS cell lines exhibited typical ES cell morphology and marker expression, but varied in their in vitro/in vivo differentiation ability. Global gene transcription analysis revealed that core pluripotency genes were expressed at lower levels in iPS cell lines compared to D3-ES cells (Pou5f1: x1.6~2.2-fold, Sox2: x2.58~10.0-fold, Eras: x1.08~2.54-fold, Dppa5a: x1.04~1.41-fold), while other genes showed higher expression in iPS cells (Lin28: x1.43~2.33-fold; Dnmt3b: x1.33~2.64-fold). This pattern was repeated in a survey of specific functional groups of genes (surface markers, cell death, JAK-STAT and P13K-AKT signaling pathways, endothelial, cardiovascular, and neurogenesis genes). Among the iPS cell lines examined, only two showed similar characteristics to ES cells. These results demonstrated that, in addition to cellular characterization, the numerical evaluation of gene expression using DNA microarrays might help to identify the stem cell stability and pluripotency of iPS cells.

Ago2 and GW182 expression in mouse preimplantation embryos: a link between microRNA biogenesis and GW182 protein synthesis.

MicroRNA-mediated RNA interference appears to play a role in early development and differentiation processes in preimplantation embryos. However, the expression of its key effectors, including Ago2, a key component of the RNA-induced silencing complex, and GW182, a critical component of GW bodies (GWBs), has not been assessed in preimplantation embryos. To characterise the roles of Ago2 and GW182 in early embryo development, we determined their transcription and protein synthesis in mouse embryos. Transcript levels of Ago2 and GW182 increased steadily from the one-cell stage through to the blastocyst stage when data were not normalised against an internal reference. However, when normalised against the internal standard, transcript levels for both genes were highest in four-cell stage embryos and decreased steadily through to the blastocyst stage. Indirect immunocytochemistry showed that both AGO2 and GW182 proteins were expressed in each stage in the early embryo and were observed to colocalise in the morula and blastocyst stages. Specific silencing of mRNA expression by short interference (si) RNA against Ago2 or Dicer1 decreased the expression of selected apoptosis- and development-related microRNAs, but did not inhibit development up to the blastocyst stage. However, transcription levels of Oct3/4, Nanog and Sox2 were decreased in both Ago2- and Dicer1-knockdown embryos at the blastocyst stage. Furthermore, although knockdown of these genes did not change transcript levels of GW182, GW182 protein synthesis was decreased in blastocyst stage embryos. These results suggest that Ago2 and Dicer1 regulate GW182 protein expression in mouse embryos, which is linked to microRNA biogenesis and likely to be important for differentiation in the blastocyst stage.

Hyperglycemia reduces mitochondrial content and glucose transporter expression in mouse embryos developing in vitro.

The objective of this research was to examine the effects of high concentrations of glucose on mouse embryos developing in vitro by studying embryo viability, mitochondrial content and expression of glucose transporters. Addition of 55 mM glucose to the culture medium of two-cell stage embryos significantly reduced the formation of morulae and blastocysts, resulting in fewer cells in the blastocyst stage embryos and increased levels of apoptosis. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression levels of the pro-apoptotic genes Bax and Casp3 at the blastocyst stage were increased significantly by the addition of either 25 or 55 mM glucose to the culture medium. However, addition of 25 or 55 mM glucose to the culture medium did not change the copy numbers of the apoptosis-related miRNAs mmu-mir-15a, mmu-mir-16 and mmu-mir-21. MitoTracker Green fluorescence revealed a decrease in the mitochondrial mass. The expression levels of the mitochondrial DNA-encoded genes Cox1 and Cox2 decreased sharply with the addition of 25 or 55 mM glucose to the culture medium. Both transcripts and protein synthesis of the glucose transporters Glut1 and Glut3 were reduced in blastocysts cultured in the presence of either 25 or 55 mM glucose. These results suggest that hyperglycemia reduces both mitochondrial content and expression levels of glucose transporters in mouse embryos developing in vitro and that this may result in apoptosis in these embryos.

A link between the interleukin-6/Stat3 anti-apoptotic pathway and microRNA-21 in preimplantation mouse embryos.

Signal transducers and activators of transcription-3 (Stat3) plays a central role in interleukin-6 (IL-6)-mediated cell proliferation by inhibiting apoptosis in a variety of cell types. MicroRNA-21 (miRNA-21), a ubiquitous miRNA, acts as an anti-apoptotic factor that seems to be indirectly but strictly linked to Stat3. In order to determine whether the IL-6 induced Stat3 anti-apoptosis pathway is linked with miRNA-21, we first determined the effects of recombinant mouse IL-6 on Stat3 expression, mouse embryo viability, and the mRNA levels of apoptosis related genes and miRNA-21 during mouse embryo development in vitro. Addition of 10 or 100 ng/ml of recombinant IL-6 to the culture medium did not affect the developmental ability of 2-cell stage embryos into blastocysts. However, total cell number was significantly increased and apoptosis was reduced in blastocyst stage embryos cultured in the presence of 100 ng/ml of recombinant IL-6. Furthermore, addition of recombinant IL-6 to the culture medium significantly increased the copy numbers of anti-apoptotic miRNA-21, up-regulated Bcl2l1, and down-regulated casp3. Similarly, the injection of mature miRNA-21 into cells up-regulated Bcl2l1 and down-regulated casp3. These results suggest that the induction of the Stat3 anti-apoptotic pathway by IL-6 is linked to miRNA-21 expression, which possibly results in the regulation of cell apoptosis in early mouse embryo development.

Leptin accelerates pronuclear formation following intracytoplasmic sperm injection of porcine oocytes: Possible role for MAP kinase inactivation.

Leptin, a multifunctional hormone, is present in mammalian oocytes and follicular fluids and cumulus cells. While leptin modulates oocyte maturation in vitro which seems to result in enhancement of embryo development, it is unclear whether leptin treatment of oocytes affects cytoplasmic maturation and fertilization processes. In order to gain a better understanding of the role of leptin during oocyte maturation, we examined microtubule and microfilament assembly following oocyte maturation and blastocyst formation, mitogen-activated protein kinase (MAPK) activity, and pronuclear formation following parthenogenetic stimuli or intracytoplasmic sperm injection (ICSI) in leptin-treated oocytes. Addition of 10 or 100 ng/ml leptin during oocyte maturation did not increase the proportion of metaphase II oocytes, but enhanced development to blastocyst stage by day 7 (P<0.01) after parthenogenetic activation (PA), accompanied by increased cell number. However there was no effect on the number of apoptotic cells in blastocysts. Following maturation in the presence of leptin, there were more oocytes with normal spindle formation. MAPK activity decreased more rapidly, and pronuclear formation was accelerated after parthenogenetic activation or ICSI of leptin-treated oocytes. These results suggested that exogeneous leptin enhanced spindle assembly and accelerated pronuclear formation following fertilization, possibly via the MAPK pathway.

Freezability of rat epididymal sperm induced by raffinose in modified Krebs-Ringer bicarbonate (mKRB) based extender solution.

The objective of this study was to develop an ideal freezing extender and method for rat epididymal sperm cryopreservation. Epididymal sperm collected from 30 Wistar males was frozen, and experiments were conducted to study its post-thaw characteristics when freezing with raffinose-free buffer or various concentrations of raffinose and egg yolk dissolved in distilled and deionised water, PBS, or modified Krebs-Ringer bicarbonate (mKRB)-based extender. Different concentrations of glycerol, Equex STM, or sodium dodecyl sulfate (SDS) dissolved in either PBS or mKRB containing egg yolk were also tested. Based on the data from these experiments, further experiments tested how different sugars such as raffinose, trehalose, lactose, fructose, and glucose dissolved in mKRB with Equex STM, SDS and egg yolk supplementation affected the post-thaw characteristics of cryopreserved sperm. Cryosurvival of frozen-thawed sperm were judged by microscopic assessment of the sperm motility index (SMI), and acrosome integrity was measured using FITC-PNA staining. Thawed sperm were subjected to 3h of a thermal resistance test. Beneficial effects on the post-thaw survival of sperm were obtained when 0.1M raffinose in mKRB was used with 0.75% Equex STM, 0.05% SDS, and 20% egg yolk. Sperm cryopreserved with this treatment exhibited a higher motility index and maintained greater SMI and acrosome integrity throughout incubation when compared to sperm frozen in various concentrations of other cryoprotectants and trehalose, lactose, fructose, glucose. In conclusion, cryopreservation in an extender solution of raffinose dissolved in mKRB containing Equex STM, SDS and egg yolk greatly enhances the freezability of rat epididymal sperm.

Trehalose enhanced the freezability of Poodle dog sperm collected by an artificial vagina (AV).

In an attempt to develop a suitable freezing method for Poodle dog sperm, an experiment was conducted to investigate semen collection methods of digital stimulation and an artificial vagina (AV), using Tris and trehalose-egg yolk extender, on the characteristics and cryopreservation of sperm. Two dogs (dogs A and B) were subjected to semen collection by digital stimulation and AV. The volume, sperm concentration, sperm motility index (SMI) and acrosome status of ejaculates were determined immediately after collection. The remainder was frozen as pellets in Tris and trehalose-egg yolk extender. Sperm motility index was evaluated after thawing and during a thermal resistance test, and acrosome integrity was also assessed. No significant differences regarding sperm concentrations, SMI and acrosome integrity were observed between semen collected by AV and digital stimulation. However, when dog sperm were collected by an AV and frozen in trehalose-egg yolk extender, the motility index of frozen-thawed sperm was significantly improved compared to sperm frozen in Tris-egg yolk extender which were collected by digital stimulation. In conclusion, semen collected by an AV and frozen in trehalose-egg yolk extender was effective in enhancing the freezability of Poodle dog sperm.

Effect of relaxin on in vitro fertilization of porcine oocytes.

Porcine relaxin is a peptide hormone belonging to the insulin super family that has a variety of biological functions. The present experiment was designed to investigate the effects of relaxin on sperm function and on in vitro fertilization (IVF) of porcine oocytes. Porcine spermatozoa were washed, swum-up, and incubated for 1-4 h in mTALP medium supplemented with 0, 20 or 50 ng/ml porcine relaxin. Motility was determined by observing the type of forward movement of the spermatozoa, and acrosome status was evaluated by applying the triple staining technique. Immature oocytes were aspirated from antral follicles and matured in IVM medium (modified NCSU-37). Matured oocytes were co-cultured with spermatozoa in IVF medium (mTALP) supplemented with 0, 5, 10, 15 or 20 ng/ml relaxin. After 6 h of sperm-oocyte co-incubation, putative zygotes were cultured for 18 h in oocyte culture medium NCSU-37 and then assessed for the rates of monospermy, polyspermy, and male pronucleus formation after acetic orcein staining. Relaxin improved (P<0.05) sperm motility and increased the percentage of acrosome-reacted live spermatozoa during 1-4 h of incubation, although viability was not significantly improved. Significantly (P<0.05) the highest percentage of monospermic (31.7%) and lowest percentage of polyspermic (16.5%) fertilization was achieved from the sperm-oocyte co-culture group treated with 20 ng/ml relaxin as compared to other groups. The percentage of male pronucleus formation was significantly (P<0.05) greater in the 20 ng/ml relaxin-treated sperm-oocyte co-culture group than in the other groups. These results indicate that supplementation with relaxin is capable of improving sperm function and fertilization of porcine oocytes in vitro.