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        체외수정 유래 생쥐 배아줄기세포와 유사한 특성을 보유한 단위발생 유래 생쥐 배아줄기세포

        박세필,김은영,이금실,이영재,신현아,민현정,이훈택,정길생,임진호,Park, Se-Pill,Kim, Eun-Young,Lee, Keum-Si,Lee, Young-Jae,Shin, Hyun-Ah,Min, Hyun-Jung,Lee, Hoon-Taek,Chung, Kil-Saeng,Lim, Jin-Ho 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.2

        Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.

      • 형광 동소 보합법을 이용한 췌장암의 세포유전학적 변화에 대한 분석과 그 임상적 의의

        윤유석(Yoo-Seok Yoon),이동순(Dong Soon Lee),민현정(Hyun Chung Min),장진영(Jin-Young Jang),이승은(Seung Eun Lee),황대욱(Dae Wook Hwang),한호성(Ho-Seong Han),김선회(Sun-Whe Kim) 한국간담췌외과학회 2008 한국간담췌외과학회지 Vol.12 No.1

        Purpose: The purposes of this study are to examine the cytogenetic alterations of pancreatic cancer, by using fluorescent in situ hybridization (FISH), to determine thier correlation with the clinico-pathologic prognostic factors and to identify the cytogenetic factors that can predict the prognosis of pancreatic cancer. Methods: Fresh frozen tissues of pancreatic cancer and normal pancreas that were obtained via pancreatic resection from 20 patients with pancreatic ductal adenocacinoma were analyzed by performing FISH with using locus-specific c-myc, p16, p53 probes and chromosome 18q, 20q probes. We cpmpared the FISH results with the clinico-pathologic prognostic factors. We also examined 16 paraffin-embedded tissues of pancreatic cancer by performing immunohistochemical staining (IHC) with monoclonal antibody to c-myc, p16, p53 and DPC. We then evaluated the correlation between the results of FISH and the results of IHC. Results: At least one alteration of genes or chromosomes was detected in 18 (90.0%) of the 20 pancreatic cancer tissues by FISH, as compared with no alternation in the normal pancreatic tissues: these alteration were an increased copy number of c-myc (66.7%), a decreased copy number of p16 (70.6%), deletion of p53 (100%), loss of chromosome 18q (56.3%) and gain of chromosome 20q (45.0%). IHC demonstrated overexpression of c-myc and p53 in 31.3% and 50.0% of the pancreatic cancer specimens, respectively, and the loss of expressions of p16 and DPC in 25.0% and 93.3% of the pancreatic cancer specimens, respectively. The concordance rate of IHC with FISH was 33.3% to 61.5%. Analysis of the correlation between the cytogenetic changes identified by FISH or IHC and the pathologic prognostic factors showed that only chromosome 20q gain was significantly correlated with the histologic grade (p=0.098) and lymphovascular invasion (p=0.092). However there was no clinical correlation of the cytogenetic changes with respect to recurrence after operation. Conclusion: This study confirms that most pancreatic cancers have cytogenetic alternations, as can be determined by FISH. Especially, the correlation between chromosome 20q gain and the prognostic pathologic factors offers the possibility of a new prognostic biologic marker located in chromosome 20q. However, further studies with more cases are needed to clarify the clinical significance of cytogenetic alternations in pancreatic cancer.

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