大连医科大学学报
大連醫科大學學報
대련의과대학학보
JOURNAL OF DALIAN MEDICAL UNIVERSITY
2015年
4期
331-335
,共5页
杜莉莉%吕润潇%于艳秋%辛娜%金玉楠
杜莉莉%呂潤瀟%于豔鞦%辛娜%金玉楠
두리리%려윤소%우염추%신나%금옥남
巨噬细胞%小鼠%迁移%细胞因子
巨噬細胞%小鼠%遷移%細胞因子
거서세포%소서%천이%세포인자
macrophages%mice%migration%cytokine
目的:比较不同方法分离的小鼠腹腔巨噬细胞的生物学特性并找出最优方法。方法采用3种方法分离小鼠腹腔巨噬细胞:5%淀粉肉汤小鼠腹腔注射3 d后灌洗(5%淀粉肉汤组);直接DMEM液腹腔灌洗(无血清DMEM组);含75%血清DMEM液腹腔注射30 min后灌洗(75%血清DMEM组),每组6只小鼠。观察各组巨噬细胞形态、计数细胞数量、流式细胞仪测细胞纯度、台盼蓝染色测细胞存活率、划痕实验测迁移能力及ELISA检测脂多糖( LPS)刺激后巨噬细胞分泌TNFα、IL-6及IL-1β的水平。结果3组巨噬细胞的形态、纯度、存活率相似,75%血清DMEM组巨噬细胞数量为(80.47±0.82)×105个,明显多于5%淀粉肉汤组(5.64±0.31)×105个及无血清DMEM 组(2.63±0.42)×105个(P<0.05);75%血清DMEM组巨噬细胞迁移能力明显高于其他两组(P<0.05);75%血清DMEM组巨噬细胞分泌TNFα(15.51±0.51)ng/mL、IL-6(415.33±1.55)pg/mL、IL-1β(421.01±2.64)pg/mL,均多于5%淀粉肉汤组TNFα(10.50±0.50)ng/mL、IL-6(386.33±1.52)pg/mL、IL-1β(375.33±2.51)pg/mL(P<0.05),也多于无血清DMEM组TNFα(9.56±0.25)ng/mL、IL-6(384.66±1.15)pg/mL、IL-1β(393.34±2.62)pg/mL(P<0.05)。结论75%血清DMEM液腹腔注射30min后灌洗是一个比较好的分离小鼠腹腔巨噬细胞的方法。
目的:比較不同方法分離的小鼠腹腔巨噬細胞的生物學特性併找齣最優方法。方法採用3種方法分離小鼠腹腔巨噬細胞:5%澱粉肉湯小鼠腹腔註射3 d後灌洗(5%澱粉肉湯組);直接DMEM液腹腔灌洗(無血清DMEM組);含75%血清DMEM液腹腔註射30 min後灌洗(75%血清DMEM組),每組6隻小鼠。觀察各組巨噬細胞形態、計數細胞數量、流式細胞儀測細胞純度、檯盼藍染色測細胞存活率、劃痕實驗測遷移能力及ELISA檢測脂多糖( LPS)刺激後巨噬細胞分泌TNFα、IL-6及IL-1β的水平。結果3組巨噬細胞的形態、純度、存活率相似,75%血清DMEM組巨噬細胞數量為(80.47±0.82)×105箇,明顯多于5%澱粉肉湯組(5.64±0.31)×105箇及無血清DMEM 組(2.63±0.42)×105箇(P<0.05);75%血清DMEM組巨噬細胞遷移能力明顯高于其他兩組(P<0.05);75%血清DMEM組巨噬細胞分泌TNFα(15.51±0.51)ng/mL、IL-6(415.33±1.55)pg/mL、IL-1β(421.01±2.64)pg/mL,均多于5%澱粉肉湯組TNFα(10.50±0.50)ng/mL、IL-6(386.33±1.52)pg/mL、IL-1β(375.33±2.51)pg/mL(P<0.05),也多于無血清DMEM組TNFα(9.56±0.25)ng/mL、IL-6(384.66±1.15)pg/mL、IL-1β(393.34±2.62)pg/mL(P<0.05)。結論75%血清DMEM液腹腔註射30min後灌洗是一箇比較好的分離小鼠腹腔巨噬細胞的方法。
목적:비교불동방법분리적소서복강거서세포적생물학특성병조출최우방법。방법채용3충방법분리소서복강거서세포:5%정분육탕소서복강주사3 d후관세(5%정분육탕조);직접DMEM액복강관세(무혈청DMEM조);함75%혈청DMEM액복강주사30 min후관세(75%혈청DMEM조),매조6지소서。관찰각조거서세포형태、계수세포수량、류식세포의측세포순도、태반람염색측세포존활솔、화흔실험측천이능력급ELISA검측지다당( LPS)자격후거서세포분비TNFα、IL-6급IL-1β적수평。결과3조거서세포적형태、순도、존활솔상사,75%혈청DMEM조거서세포수량위(80.47±0.82)×105개,명현다우5%정분육탕조(5.64±0.31)×105개급무혈청DMEM 조(2.63±0.42)×105개(P<0.05);75%혈청DMEM조거서세포천이능력명현고우기타량조(P<0.05);75%혈청DMEM조거서세포분비TNFα(15.51±0.51)ng/mL、IL-6(415.33±1.55)pg/mL、IL-1β(421.01±2.64)pg/mL,균다우5%정분육탕조TNFα(10.50±0.50)ng/mL、IL-6(386.33±1.52)pg/mL、IL-1β(375.33±2.51)pg/mL(P<0.05),야다우무혈청DMEM조TNFα(9.56±0.25)ng/mL、IL-6(384.66±1.15)pg/mL、IL-1β(393.34±2.62)pg/mL(P<0.05)。결론75%혈청DMEM액복강주사30min후관세시일개비교호적분리소서복강거서세포적방법。
Objective To compare the characteristics of mouse peritoneal macrophages isolated by different methods and find out the best method.Methods Murine peritoneal macrophages were isolated by three methods:intraperitoneal injection of 5%starch broth for 3 d then lavage(5%starch broth group);direct DMEM solution lavage ( non serum DMEM group);intraperitoneal injection of DMEM solution containing 75%serum for 30 min then lavage (75%serum DMEM group).Re-sults The morphology, purity and survival rate of macrophages in three groups were similar., the number of macrophages in 75%serum DMEM group was (80.47 ±0.82) ×105 , significantly more than 5%starch broth group(5.64 ±0.31) ×105 and non serum DMEM group(2.63 ±0.42) ×105(P<0.05), the migration ability of macrophages in 75%serum DMEM group was significantly higher than those in other two groups (P<0.05).Macrophages in 75%serum DMEM group secre-ted TNFα(15.51 ±0.51) ng/mL, IL-6(415.33 ±1.55) pg/mL and IL-1β(421.01 ±2.64) pg/mL, more than 5%starch broth group TNFα(10.50 ±0.50) ng/mL, IL-6 (386.33 ±1.52) pg /mL and IL-1β(375.33 ±2.51) pg/mL (P<0.05), also more than non serum DMEM group TNFα(9.56 ±0.25) ng/mL, IL-6 (384.66 ±1.15) pg/mL, IL-1β(393.34 ±2.62)pg/mL (P<0.05).Conclusion Intraperitoneal injection of 75% serum DMEM for 30 min then lav-age, is a good method for extraction of mouse peritoneal macrophages.