畜牧与饲料科学
畜牧與飼料科學
축목여사료과학
ANIMAL HUSBANDRY AND FEED SCIENCE
2011年
9期
96-99
,共4页
杨苗%考桂兰%侯先志%高爱武%李大彪%王秀美%杨银芬
楊苗%攷桂蘭%侯先誌%高愛武%李大彪%王秀美%楊銀芬
양묘%고계란%후선지%고애무%리대표%왕수미%양은분
奶牛%乳腺上皮细胞%冻存液
奶牛%乳腺上皮細胞%凍存液
내우%유선상피세포%동존액
dairy cows%mammary epithelial cells%cryoprotectants
试验旨在探究奶牛乳腺上皮细胞（bovine mammary epithelial cells,BMECs）最佳的冻存液以改善乳腺上皮细胞的冻存质量。BMECs传至第5代后分别加入以下10种不同配方的冻存液。A组：85%DMEM＋10%胎牛血清＋5%DMSO;B组：80%DMEM＋10%胎牛血清＋10%DMSO;C组：75%DMEM＋10%胎牛血清＋15%DMSO;D组：70%DMEM＋10%胎牛血清＋20%DMSO;E组：85%DMEM＋5%胎牛血清＋10%DMSO;F组：75%DMEM＋15%胎牛血清＋10%DMSO;G组：70%DMEM＋20%胎牛血清＋10%DMSO;H组：80%DMEM＋10%胎牛血清＋10%甘油;I组：70%DMEM＋10%胎牛血清＋20%甘油;J组：60%DMEM＋10%胎牛血清＋30%甘油,冻存前统一调整细胞密度到1×106个/mL冻存。分别对复苏后的细胞进行台盼蓝染色计算存活率和PI/Hoechst33258双染计算凋亡率。结果表明,BMECs经不同冻存剂冻存复苏后,细胞活力、形态学及凋亡率表现有所不同,其中B组和G组的活力和24h贴壁率较其他组高,二者的凋亡率较低,二者之间差异无显著性（P〉0.05）;传代后B组细胞的生长状况最好。
試驗旨在探究奶牛乳腺上皮細胞（bovine mammary epithelial cells,BMECs）最佳的凍存液以改善乳腺上皮細胞的凍存質量。BMECs傳至第5代後分彆加入以下10種不同配方的凍存液。A組：85%DMEM＋10%胎牛血清＋5%DMSO;B組：80%DMEM＋10%胎牛血清＋10%DMSO;C組：75%DMEM＋10%胎牛血清＋15%DMSO;D組：70%DMEM＋10%胎牛血清＋20%DMSO;E組：85%DMEM＋5%胎牛血清＋10%DMSO;F組：75%DMEM＋15%胎牛血清＋10%DMSO;G組：70%DMEM＋20%胎牛血清＋10%DMSO;H組：80%DMEM＋10%胎牛血清＋10%甘油;I組：70%DMEM＋10%胎牛血清＋20%甘油;J組：60%DMEM＋10%胎牛血清＋30%甘油,凍存前統一調整細胞密度到1×106箇/mL凍存。分彆對複囌後的細胞進行檯盼藍染色計算存活率和PI/Hoechst33258雙染計算凋亡率。結果錶明,BMECs經不同凍存劑凍存複囌後,細胞活力、形態學及凋亡率錶現有所不同,其中B組和G組的活力和24h貼壁率較其他組高,二者的凋亡率較低,二者之間差異無顯著性（P〉0.05）;傳代後B組細胞的生長狀況最好。
시험지재탐구내우유선상피세포（bovine mammary epithelial cells,BMECs）최가적동존액이개선유선상피세포적동존질량。BMECs전지제5대후분별가입이하10충불동배방적동존액。A조：85%DMEM＋10%태우혈청＋5%DMSO;B조：80%DMEM＋10%태우혈청＋10%DMSO;C조：75%DMEM＋10%태우혈청＋15%DMSO;D조：70%DMEM＋10%태우혈청＋20%DMSO;E조：85%DMEM＋5%태우혈청＋10%DMSO;F조：75%DMEM＋15%태우혈청＋10%DMSO;G조：70%DMEM＋20%태우혈청＋10%DMSO;H조：80%DMEM＋10%태우혈청＋10%감유;I조：70%DMEM＋10%태우혈청＋20%감유;J조：60%DMEM＋10%태우혈청＋30%감유,동존전통일조정세포밀도도1×106개/mL동존。분별대복소후적세포진행태반람염색계산존활솔화PI/Hoechst33258쌍염계산조망솔。결과표명,BMECs경불동동존제동존복소후,세포활력、형태학급조망솔표현유소불동,기중B조화G조적활력화24h첩벽솔교기타조고,이자적조망솔교저,이자지간차이무현저성（P〉0.05）;전대후B조세포적생장상황최호。
This paper was aimed to search an optimal cryoprotectant for improving the cryopreservation quality of bovine mammary epithelial cells.The following 10 different ingredients of cryoprotectant were added to the fifth generation of BMECs respectively,group A：85% DMEM＋10% FBS＋5% DMSO;group B：80% DMEM＋10% FBS＋10% DMSO;group C：75% DMEM＋10% FBS＋15% DMSO;group D：70% DMEM＋10% FBS＋20% DMSO;group E：85% DMEM＋5% FBS＋10% DMSO;group F：75% DMEM＋15% FBS＋10% DMSO;group G：70% DMEM＋20% FBS＋10% DMSO;group H：80% DMEM＋10% FBS＋10%glycerol;group I：70% DMEM＋10% FBS＋20% glycerol;group J：60% DMEM＋10% FBS＋30% glycerol.The cell density was adjusted to 1×106/mL before cryopreservation.After the cell recovery,the survival and apoptosis rate of cells were calculated by trypan blue staining and PI/Hoechst 33258 double staining respectively.The results indicated that the BMECs in these groups showed different levels of viability,morphology,and apoptosis after recovery.The vitality and 24 h adhering rate of group B and G was higher than that in other groups.Furthermore,the apoptosis rate of this two groups was lower and there were no significant difference between them （P0.05）.After passage,the growth state of BMECs was the best for group B.
