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SCI논문 번역에 대해서 알아 보겠습니다(한영번역)

SCI논문 번역(한국어 원본)

24well plate에 HA, BGS-7를 끼워 넣고 시편과 plate에 BMSC를 2x104cells/well되도록 seeding한 후 다음날 분화 media에 alendronate를 농도가 10-5 -10-10 M이 되도록 첨가하였다. 1주, 3주 후에 each well was washed in 1X Hank’s Buffer (Sigma-Aldrich, Brondby, Denmark). And 500ul Alkaline Buffer (Sigma-Aldrich, Brondby, Denmark) was added to each well and these wells were shaking cultured for 1 hour at room temperature. After this period, 10ul of lysates was added to each well in a 96-well plate, and then dilution procedure was performed by adding 50ul Alkaline Buffer. Upon the completion of this procedure, 50ul of 2-amino-2-methyl-1-propanol buffer (Sigma-Aldrich, Brondby, Denmark) was added, followed by 4 hours of storing at room temperature. The determination of ALP activity is based on the ability of this enzyme to hydrolyze the paranitrophenyl phosphate in an end-point assay in which yellow product paranitrophenol (p-nitrophenol) is produced. The rate of p-nitrophenol product is proportional to the amount of ALP in the solution. The ALP activity in the culture media was read at 405 nm by ELISA Reader. ALP assay는 triplicate로 진행되었다. ALP staining을 위하여 1주와 3주 후에 cell을 고정 시키고 각 well에 Fast blue RR salt와 naphtol AS-MX phosphate alkaline solution 500ul씩 첨가하여 빛을 차단한 상태로 room temperature에서 1시간 동안 incubation후 water로 washing을 하였다.
3) RT-PCR
24well plate에 HA, BGS-7를 끼워 넣고 시편과 plate에 BMSC를 2x104cells/well되도록 seeding한 후 다음날 분화 media에 alendronate를 농도가 10-7 M이 되도록 첨가하였다. 1주, 3주 후에 각각 easy-blue RNA isolation kit(Intron사)를 이용하여 total RNA를 얻었다. Nano drop을 이용하여 total RNA를 정량하고 RNA 2ug으로 superscript III(Invitrogen사)를 이용하여 cDNA를 합성하였다 (Table 1). cDNA합성 후 ALP, osteopontin, osteocalin, runx2, GAPDH에 대하여 gene specific PCR을 진행하였다. 모든 PCR products는 1.5% agarose gel에 가시화 되고 gel doc에 의하여 상대적인 gene expression이 정성분석 하였다.

SCI논문 번역(영어 번역본)

HA and BGS-7 were inserted into a well of 24-well plate and BMSCs were seeded into the samples and the plate at a density of 2x104cells/well. Next day, alendronate at a concentration of between 10-5 and -10-10M was added to the culture media. At the end of week 1 and week 3, each well was washed in 1X Hank’s Buffer (Sigma-Aldrich, Brondby, Denmark). And 500ul Alkaline Buffer (Sigma-Aldrich, Brondby, Denmark) was added to each well and these wells were shaking cultured for 1 hour at room temperature. After this period, 10ul of lysates was added to each well in a 96-well plate, and then dilution procedure was performed by adding 50ul Alkaline Buffer. Upon the completion of this procedure, 50ul of 2-amino-2-methyl-1-propanol buffer (Sigma-Aldrich, Brondby, Denmark) was added, followed by 4 hours of storing at room temperature. The determination of ALP activity is based on the ability of this enzyme to hydrolyze the paranitrophenyl phosphate in an end-point assay in which yellow product paranitrophenol (p-nitrophenol) is produced. The rate of p-nitrophenol product is proportional to the amount of ALP in the solution. The ALP activity in the culture media was read at 405 nm by ELISA Reader. The ALP assay was carried out in triplicate. For the ALP staining, the cells were fixed at the end of week 1 and week 3, and Fast blue RR salt and naphtol AS-MX phosphate alkaline solution (500ul each) were added to each well. And then the cells were incubated for one hour at room temperature with all lights blocked and were washed in water.
3) RT-PCR
HA and BGS-7 were inserted into a well of 24-well plate and BMSCs were seeded into the samples and the plate at a density of 2x104cells/well. Next day, alendronate at a concentration of 10-7M was added to the culture media. At the end of week 1 and week 3, total RNA was obtained using easy-blue RNA isolation kit (Intron Co., Inc.). Total RNA was quantified using nano drop and cDNA was synthesized with 2ug of RNA using superscript III (Invitrogen Co., Inc.). (Table 1) After cDNA synthesis, gene specific PCR was carried out on ALP, osteopontin, osteocalcin, runx2 and GAPDH. All PCR products were visualized on 1.5% agarose gel and qualitative analysis was performed on the relative gene expression by gel doc.

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