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您現在的位置:首頁  >  技術文章  >  C2C12 鼠肌原(yuan)細胞, 進口胎(tai)牛血清(qing)培(pei)養(yang)

C2C12 鼠肌原細胞, 進口胎牛血清培養

發布時間:2016-05-04瀏覽(lan):1480次

C2C12 鼠肌原細胞, 進口胎牛(niu)血清培養慧穎生物一站式采購商(shang)城,細(xi)胞株(zhu)細(xi)胞系,耐(nai)藥株(zhu),STR鑒(jian)定,各品牌進(jin)口(kou)胎牛血清,培養基,胰酶,雙抗,*培養基,PBS緩沖液,DMSO,細(xi)胞生長因子等,種類齊全,*!

細胞(bao)培養技(ji)術屬于細胞(bao)工程(cheng)范疇,細胞(bao)工程(cheng)包括細胞(bao)融合技(ji)術、單克(ke)隆(long)抗(kang)體(ti)的(de)制備、核(he)移植技(ji)術(克(ke)隆(long))等,而這些(xie)技(ji)術中都有細胞(bao)培養過程(cheng),所以(yi)說細胞(bao)培養是生物學研究的(de)zui基本(ben)技(ji)術之一。

上海(hai)慧穎生物供應C2C12 鼠肌(ji)原(yuan)細胞, 進口(kou)胎牛血清培(pei)養,活性好、存活率(lv)高。隨(sui)著細(xi)胞(bao)(bao)生物學尤其是干細(xi)胞(bao)(bao)研究(jiu)的(de)快(kuai)速發展(zhan),在細(xi)胞(bao)(bao)治療(liao)領域已(yi)經(jing)進行了(le)(le)大量的(de)探(tan)索,取得(de)了(le)(le)一系列的(de)令人(ren)振奮的(de)成果,且部分基礎研究(jiu)成果已(yi)經(jing)在心血管系統(tong)疾病 、神經(jing)系統(tong)疾病、肌肉(rou)骨骼(ge)相關疾病、糖尿病等。

養(yang)細(xi)胞經驗(yan)分享:

1.細(xi)(xi)(xi)胞形態觀(guan)察。細(xi)(xi)(xi)胞膜是(shi)每天(tian)必須(xu)觀(guan)察的,看是(shi)否清晰,細(xi)(xi)(xi)胞的死亡往往是(shi)從細(xi)(xi)(xi)胞膜的破裂開始的,早(zao)期的表現就(jiu)是(shi)細(xi)(xi)(xi)胞膜某(mou)一點出(chu)現欠完整或者(zhe)毛發影(ying)。

2.細胞往(wang)(wang)往(wang)(wang)會(hui)有“抱(bao)團”現(xian)象,抱(bao)團生長或(huo)死亡,個人(ren)認為應該勤換液(ye)或(huo)傳代,盡量(liang)避 免細胞抱(bao)團。

3.細(xi)胞(bao)(bao)生長迅(xun)速,強(qiang)烈建(jian)議用較大的(de)培養瓶養,用小瓶養的(de)話,換液不及時,很容易死 亡。細(xi)胞(bao)(bao)的(de)生長速度一(yi)般為2天左(zuo)右就需要傳(chuan)代,并且一(yi)般為一(yi)傳(chuan)三,或一(yi)傳(chuan)四。

4.培(pei)養液:1640|DMEM|MEM|IMDM|a-MEM培(pei)養液,1L一般加碳酸氫鈉(na)2g,并加雙抗(kang),胎牛血清濃度(du)*為12%,zui低不低于10%,有些細(xi)(xi)胞(bao)對胎牛血清高(gao)度(du)依賴,比如WRO細(xi)(xi)胞(bao),HT-22細(xi)(xi)胞(bao),HA-VSMC細(xi)胞,NK-92細(xi)胞等。培養液PH應調為7.20-7.40之間。

5.防污染。L-02細胞(bao)生長(chang)迅速,較少發生污染,但常規措施應該做好(hao)。比如無菌操(cao)作,酒精 消毒等,培養(yang)瓶口(kou)建議過(guo)火(huo),包括(kuo)瓶蓋,應過(guo)火(huo)4秒左右。

6.凍存(cun)(cun)。-20一小時,-80次夜,液(ye)氮(dan)長期保存(cun)(cun)。強烈(lie)建議液(ye)氮(dan)長期保存(cun)(cun),盡量不要在-80長期 保存(cun)(cun),死亡率很高。

7.復蘇。調節水浴(yu)箱溫度為42℃,并(bing)提前在試(shi)(shi)管(guan)內(nei)(nei)(nei)放置(zhi)10倍培養液(ye)(ye)(ye)(ye),使培養液(ye)(ye)(ye)(ye)溫度與室溫一 致。1分(fen)鐘內(nei)(nei)(nei)融化細胞(禁止搖晃凍存管(guan)),移取至試(shi)(shi)管(guan)內(nei)(nei)(nei),動作要(yao)輕,從試(shi)(shi)管(guan)管(guan)底開(kai)始慢慢上提 移液(ye)(ye)(ye)(ye)管(guan),使細胞在培養液(ye)(ye)(ye)(ye)內(nei)(nei)(nei)分(fen)布均(jun)勻,離心,洗(xi)2次。

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其它經驗概括(kuo):

細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)類型(xing)(xing)(xing)(xing)(xing) 編輯(ji) 細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)類型(xing)(xing)(xing)(xing)(xing)根(gen)據是否附(fu)(fu)于支持(chi)物上(shang)(shang)(shang)生(sheng)(sheng)(sheng)長(chang)(chang)(chang)的(de)(de)(de)特性(xing)(xing),分(fen)為 貼(tie)附(fu)(fu)型(xing)(xing)(xing)(xing)(xing)、懸(xuan)浮(fu)型(xing)(xing)(xing)(xing)(xing)。 貼(tie)附(fu)(fu)型(xing)(xing)(xing)(xing)(xing) 細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)貼(tie)附(fu)(fu)在(zai)(zai)(zai)支持(chi)物表面生(sheng)(sheng)(sheng)長(chang)(chang)(chang),只依賴貼(tie)附(fu)(fu)才能(neng)生(sheng)(sheng)(sheng)長(chang)(chang)(chang)的(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)叫做貼(tie)附(fu)(fu)型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(Anchorrage-dependent cells)這(zhe)種現象(xiang)與細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)分(fen)化有關(guan)。 貼(tie)附(fu)(fu)型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)的(de)(de)(de)分(fen)型(xing)(xing)(xing)(xing)(xing) 1.成(cheng)纖維型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao):(fibroblast) 來自(zi)中(zhong)(zhong)(zhong)胚(pei)層 特點(dian):與體內(nei)成(cheng)纖維細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)形(xing)(xing)態(tai)(tai)相似,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)體梭(suo)型(xing)(xing)(xing)(xing)(xing)或(huo)(huo)不(bu)(bu)(bu)規則(ze)三角形(xing)(xing),中(zhong)(zhong)(zhong)央有圓形(xing)(xing)核,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)質向外(wai)伸出2~3個(ge)長(chang)(chang)(chang)短不(bu)(bu)(bu)同的(de)(de)(de)突起。細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)生(sheng)(sheng)(sheng)長(chang)(chang)(chang)時(shi)呈(cheng)(cheng)放射狀(zhuang)(zhuang),漩渦(wo)或(huo)(huo)火焰狀(zhuang)(zhuang)走(zou)(zou)行(xing)。 起源:細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)來自(zi)中(zhong)(zhong)(zhong)胚(pei)層間(jian)充質組(zu)織。 除真正的(de)(de)(de)成(cheng)纖維細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、心肌(ji)、平滑(hua)肌(ji)、成(cheng)骨細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、血管內(nei)皮細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)。 注(zhu)意(yi):細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)培養時(shi)成(cheng)為成(cheng)纖維型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)是一種慣稱,與體內(nei)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)不(bu)(bu)(bu)同。 2.上(shang)(shang)(shang)皮型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(epithlium cell type) 來自(zi)外(wai)胚(pei)層。 特點(dian):扁平不(bu)(bu)(bu)規則(ze)多(duo)角型(xing)(xing)(xing)(xing)(xing)、圓形(xing)(xing)核、細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)緊密相連成(cheng)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)增殖數目(mu)增多(duo)時(shi),整個(ge)上(shang)(shang)(shang)皮膜隨之移動(dong)。邊緣細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)很(hen)少脫(tuo)離細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)群而單獨活動(dong)“ 拉網(wang)”現象(xiang)與起源內(nei)外(wai)胚(pei)層組(zu)織有關(guan)。 組(zu)織: 皮膚表皮及其衍生(sheng)(sheng)(sheng)物(汗腺、皮脂腺)消化道(dao)分(fen)上(shang)(shang)(shang)皮、肝、胰、肺泡上(shang)(shang)(shang)皮。 3. 游(you)走(zou)(zou)型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(wandering cell type) 特點(dian):在(zai)(zai)(zai)支持(chi)物上(shang)(shang)(shang)散在(zai)(zai)(zai)生(sheng)(sheng)(sheng)長(chang)(chang)(chang),不(bu)(bu)(bu)連接(jie)(jie)成(cheng)片,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)質伸出偽足或(huo)(huo)突起呈(cheng)(cheng)活躍的(de)(de)(de)游(you)走(zou)(zou)或(huo)(huo)變形(xing)(xing)運動(dong),速度快不(bu)(bu)(bu)規則(ze), 密度大連接(jie)(jie)成(cheng)片呈(cheng)(cheng)多(duo)角形(xing)(xing)不(bu)(bu)(bu)易與其他(ta)類型(xing)(xing)(xing)(xing)(xing)的(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao) 區別(bie)。 4. 多(duo)形(xing)(xing)型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(polymorphic cell type) 特點(dian): 無規律(lv)的(de)(de)(de)形(xing)(xing)態(tai)(tai),如神經細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)。 懸(xuan)浮(fu)型(xing)(xing)(xing)(xing)(xing) 特點(dian): 不(bu)(bu)(bu)貼(tie)附(fu)(fu)在(zai)(zai)(zai)支持(chi)物生(sheng)(sheng)(sheng)長(chang)(chang)(chang),胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)體圓形(xing)(xing),在(zai)(zai)(zai)培養液中(zhong)(zhong)(zhong)生(sheng)(sheng)(sheng)長(chang)(chang)(chang)空(kong)間(jian)大,可(ke)長(chang)(chang)(chang)時(shi)間(jian)的(de)(de)(de)生(sheng)(sheng)(sheng)長(chang)(chang)(chang),繁(fan)殖旺盛(sheng)便于做細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)代謝研究。S180肉瘤、血液里的(de)(de)(de)白細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、K562、 HL-60。 分(fen)型(xing)(xing)(xing)(xing)(xing)的(de)(de)(de)目(mu)的(de)(de)(de): 方便描(miao)述細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)培養過(guo)程中(zhong)(zhong)(zhong)的(de)(de)(de)形(xing)(xing)態(tai)(tai)變化。培養條件好時(shi),細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)相對穩定,可(ke)反映出起源、正常異常的(de)(de)(de)區別(bie),作為判定細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)生(sheng)(sheng)(sheng)物學(xue)性(xing)(xing)狀(zhuang)(zhuang)指(zhi)(zhi)標。 強調:一般(ban)形(xing)(xing)態(tai)(tai)并不(bu)(bu)(bu)是一項(xiang)可(ke)靠指(zhi)(zhi)標要受(shou)各方面影(ying)響。 如:反復(fu)開關(guan)溫箱、溫度、C02的(de)(de)(de)濃度、培養基變堿、支原體、pH值。細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)接(jie)(jie)種時(shi)呈(cheng)(cheng)三角型(xing)(xing)(xing)(xing)(xing)狀(zhuang)(zhuang)態(tai)(tai),經培養一定時(shi)間(jian)后,細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)傳(chuan)代前已(yi)形(xing)(xing)成(cheng)上(shang)(shang)(shang)皮型(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)。

KG-1a細(xi)胞,人急(ji)性骨髓(sui)白血(xue)病細(xi)胞      25T

MV4-11細(xi)胞,人急性白血病細(xi)胞    25T

HL-60細胞,人急(ji)性早幼粒(li)白血病細胞    25T

U937細(xi)胞,人急性早幼粒白血(xue)病細(xi)胞     25T

MOLM-13細胞,人急性髓(sui)系白血病細胞    25T

MOLM-14細胞,人急(ji)性髓系白血病細胞    25T

K562細胞,人慢性髓系白血病細胞  25T

HL-60細胞,人急性早幼(you)粒(li)白(bai)血病細胞    25T

CCRF-CEM細胞(bao),人急性淋巴(ba)細胞(bao)白(bai)血病T淋巴(ba)細胞(bao)  25T

HS 505.T細(xi)胞(bao),慢性淋(lin)巴細(xi)胞(bao)白(bai)血病(CLL)細(xi)胞(bao)   25T

 

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