국내 유통 중인 콜라겐 제품 120건을 대상으로 중금속 4종(납, 카드뮴, 비소, 수은) 함량에 대해 조사하였다. 수 은은 금아말감화법을 이용한 수은분석기로 분석하였고 납, 카드뮴, 비소는 ICP-OES를 이용하여 분석하였다. 검사 결 과 납의 평균 함량은 0.097±0.055 mg/kg이었고 건강기능 식품, 기타가공품, 음료류, 과·채가공품에서 각각 평균 0.108±0.052mg/kg, 0.084±0.053 mg/kg, 0.131±0.047 mg/ kg, 0.149 mg/kg 농도로 검출되었다. 카드뮴의 평균 함량 은 0.026±0.011 mg/kg이었으며 검출된 제품은 모두 건강 기능식품이었다. 비소의 평균 함량은 0.097±0.048 mg/kg 이었고 건강기능식품, 기타가공품, 과·채가공품에서 각각 평균 0.091±0.055 mg/kg, 0.133 mg/kg, 0.086 mg/kg 농도 로 검출되었다. 수은의 평균 함량은 0.0025±0.0016 mg/kg 으로 건강기능식품, 기타가공품, 과·채가공품, 캔디류에서 각각 평균 0.0012 mg/kg, 0.0028±0.0018 mg/kg, 0.0013 mg/kg, 0.0031 mg/kg 농도로 검출되었다. 중금속 기준·규 격이 있는 음료류(납 0.3 mg/kg, 카드뮴 0.1 mg/kg) 및 캔 디류(납 0.2 mg/kg)는 모두 기준 이하로 검출되어 적합하 였다. 기준·규격이 없는 제품도 국내외 식품의 중금속 기 준과 보고된 식품 중금속 함량과 비교하였을 때 비교적 안전한 수준이라고 판단되었다.

본 연구에서는 chlorogenic acid 이성질체 9종과 arbutin을 분석하기 위하여, 전처리 과정과 UHPLC-MS/MS 동시 분석 조건을 확립하였다. UHPLC-MS/MS에서 C18 column을 사용하여 15분 동안 분석할 수 있는 용매 조성을 만들고, 10가지 물질에 대한 정량, 정성 이온을 선택하여 negative 모드에서 분석하는 동시분석법을 확립하였고 과일류를 대상으로 추출, 진탕 추출, 초음파 추출, 원심분리, 농축 등의 과정을 거치는 전처리법을 개발하였다. 기기분석과 전처리법에 대한 유효성은 특이성, 직선성, 정확성, 정밀성 그리고 검출한계 및 정량한계 등을 통하여 확인하였다. 회수율은 48.1-120.3%, intraday precision (RSD)는 0.4-7.9%, interday precision (RSD)는 0.0-7.2%로 확인되었다. 따라서, 본 연구에서 확립한 동시분석방법은 과일류 중 chlorogenic acid와 arbutin을 효과적으로 분석하는데 유용할 것으로 사료된다.

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of Tuj1 increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous Igf2 may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells (1.09 × 105 eell₃/ml). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% (30 × 104 eell₃/ml) or 20% KSR (4.8 × 104 cells/ml) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.