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您現在的位置:首頁  >  技術文章  >  H358小細(xi)胞肺癌,進口胎(tai)牛(niu)血清(qing)培(pei)養

H358小細胞肺癌,進口胎牛血清培養

發布時(shi)間:2016-05-04瀏(liu)覽:1496次

H358小細(xi)胞肺癌,進口(kou)胎牛血清培養慧穎生物一(yi)站式采購商城(cheng),細胞(bao)株細胞(bao)系,耐藥株,STR鑒定,各品(pin)牌進口胎(tai)牛血清(qing),培(pei)養基,胰(yi)酶,雙(shuang)抗,*培(pei)養基,PBS緩(huan)沖液(ye),DMSO,細胞(bao)生長因子等,種(zhong)類齊全,*!

細胞培(pei)(pei)養(yang)技(ji)(ji)(ji)術(shu)屬于(yu)細胞工程范疇,細胞工程包括細胞融合技(ji)(ji)(ji)術(shu)、單克隆(long)抗體的(de)制(zhi)備、核移(yi)植(zhi)技(ji)(ji)(ji)術(shu)(克隆(long))等,而這些技(ji)(ji)(ji)術(shu)中(zhong)都有細胞培(pei)(pei)養(yang)過程,所以說細胞培(pei)(pei)養(yang)是生物學研究的(de)zui基本技(ji)(ji)(ji)術(shu)之一。

上海慧穎生物(wu)供(gong)應H358小細胞(bao)肺癌,進口胎牛(niu)血清培養,活性(xing)好、存(cun)活率高。隨著細胞(bao)生物學尤其是干細胞(bao)研(yan)究的快速發展,在(zai)細胞(bao)治療領域已經(jing)(jing)進行了大量的探索,取得了一系列的令人振奮的成果(guo),且部分(fen)基礎研(yan)究成果(guo)已經(jing)(jing)在(zai)心(xin)血管系統疾(ji)(ji)病(bing) 、神經(jing)(jing)系統疾(ji)(ji)病(bing)、肌肉骨骼相關疾(ji)(ji)病(bing)、糖尿病(bing)等(deng)。

養細(xi)胞(bao)經(jing)驗(yan)分享:

1.細(xi)胞形態觀察。細(xi)胞膜(mo)是(shi)每(mei)天必須(xu)觀察的(de),看是(shi)否清(qing)晰(xi),細(xi)胞的(de)死亡往(wang)往(wang)是(shi)從細(xi)胞膜(mo)的(de)破(po)裂開始(shi)的(de),早期的(de)表現就是(shi)細(xi)胞膜(mo)某一點出現欠(qian)完整或者(zhe)毛發影。

2.細(xi)(xi)胞往往會(hui)有(you)“抱團(tuan)”現象(xiang),抱團(tuan)生(sheng)長或死亡(wang),個(ge)人認為(wei)應該勤換液或傳代(dai),盡量避 免細(xi)(xi)胞抱團(tuan)。

3.細胞生長迅(xun)速,強烈建(jian)議用較大的(de)(de)培養瓶(ping)養,用小瓶(ping)養的(de)(de)話,換(huan)液不及時,很容易死 亡。細胞的(de)(de)生長速度一般為(wei)2天左右(you)就需要傳代,并且一般為(wei)一傳三(san),或一傳四。

4.培養液:1640|DMEM|MEM|IMDM|a-MEM培養液,1L一般(ban)加(jia)碳酸氫(qing)鈉2g,并加(jia)雙(shuang)抗,胎(tai)牛血清濃度*為12%,zui低不低于10%,有些細胞對胎(tai)牛血清高(gao)度依賴(lai),比如WRO細胞,HT-22細胞,HA-VSMC細胞,NK-92細胞等。培養液PH應調為7.20-7.40之間(jian)。

5.防污(wu)染。L-02細胞生長迅速,較(jiao)少發生污(wu)染,但常規措施(shi)應(ying)該做(zuo)好。比(bi)如無菌操作,酒精 消(xiao)毒(du)等,培養(yang)瓶(ping)口建議過(guo)火,包括(kuo)瓶(ping)蓋(gai),應(ying)過(guo)火4秒左右。

6.凍存。-20一小時,-80次夜,液氮長期(qi)保存。強烈建議液氮長期(qi)保存,盡量(liang)不要在-80長期(qi) 保存,死亡(wang)率很高。

7.復蘇。調節水浴箱溫度為42℃,并提前在(zai)試管(guan)(guan)(guan)內放置10倍(bei)培(pei)養(yang)液(ye)(ye),使(shi)培(pei)養(yang)液(ye)(ye)溫度與室溫一 致(zhi)。1分鐘(zhong)內融化(hua)細胞(禁止(zhi)搖晃凍存(cun)管(guan)(guan)(guan)),移(yi)取至試管(guan)(guan)(guan)內,動作要輕,從試管(guan)(guan)(guan)管(guan)(guan)(guan)底開始(shi)慢(man)慢(man)上提 移(yi)液(ye)(ye)管(guan)(guan)(guan),使(shi)細胞在(zai)培(pei)養(yang)液(ye)(ye)內分布均勻,離心,洗2次。

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其它經(jing)驗概括:

細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)類型(xing) 編輯(ji) 細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)類型(xing)根據是(shi)否附(fu)(fu)于支(zhi)持(chi)(chi)物(wu)上生(sheng)(sheng)(sheng)(sheng)長(chang)的(de)(de)(de)(de)特(te)(te)性,分(fen)為 貼附(fu)(fu)型(xing)、懸浮型(xing)。 貼附(fu)(fu)型(xing) 細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)貼附(fu)(fu)在(zai)(zai)(zai)支(zhi)持(chi)(chi)物(wu)表面(mian)生(sheng)(sheng)(sheng)(sheng)長(chang),只依賴貼附(fu)(fu)才能生(sheng)(sheng)(sheng)(sheng)長(chang)的(de)(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)叫做(zuo)貼附(fu)(fu)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(Anchorrage-dependent cells)這種現象與(yu)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)分(fen)化(hua)有關。 貼附(fu)(fu)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)的(de)(de)(de)(de)分(fen)型(xing) 1.成(cheng)(cheng)纖(xian)維(wei)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao):(fibroblast) 來自中(zhong)胚(pei)(pei)層(ceng) 特(te)(te)點(dian)(dian):與(yu)體(ti)(ti)內(nei)成(cheng)(cheng)纖(xian)維(wei)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)形(xing)(xing)(xing)(xing)態(tai)相似,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)體(ti)(ti)梭型(xing)或(huo)(huo)不(bu)(bu)規則(ze)(ze)三(san)角(jiao)形(xing)(xing)(xing)(xing),中(zhong)央(yang)有圓形(xing)(xing)(xing)(xing)核,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)質向外(wai)伸出(chu)2~3個長(chang)短不(bu)(bu)同(tong)的(de)(de)(de)(de)突起(qi)。細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)生(sheng)(sheng)(sheng)(sheng)長(chang)時(shi)呈放射(she)狀(zhuang),漩(xuan)渦或(huo)(huo)火焰狀(zhuang)走行。 起(qi)源:細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)來自中(zhong)胚(pei)(pei)層(ceng)間充質組織(zhi)。 除真正的(de)(de)(de)(de)成(cheng)(cheng)纖(xian)維(wei)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、心肌、平滑肌、成(cheng)(cheng)骨(gu)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、血管(guan)內(nei)皮(pi)(pi)(pi)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)。 注意:細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)培(pei)(pei)養(yang)(yang)時(shi)成(cheng)(cheng)為成(cheng)(cheng)纖(xian)維(wei)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)是(shi)一(yi)種慣稱,與(yu)體(ti)(ti)內(nei)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)不(bu)(bu)同(tong)。 2.上皮(pi)(pi)(pi)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(epithlium cell type) 來自外(wai)胚(pei)(pei)層(ceng)。 特(te)(te)點(dian)(dian):扁平不(bu)(bu)規則(ze)(ze)多角(jiao)型(xing)、圓形(xing)(xing)(xing)(xing)核、細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)緊密相連成(cheng)(cheng)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)增殖數目增多時(shi),整個上皮(pi)(pi)(pi)膜隨之移動。邊緣(yuan)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)很(hen)少脫離細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)群(qun)而單獨活動“ 拉網”現象與(yu)起(qi)源內(nei)外(wai)胚(pei)(pei)層(ceng)組織(zhi)有關。 組織(zhi): 皮(pi)(pi)(pi)膚表皮(pi)(pi)(pi)及其(qi)衍生(sheng)(sheng)(sheng)(sheng)物(wu)(汗腺(xian)、皮(pi)(pi)(pi)脂腺(xian))消(xiao)化(hua)道分(fen)上皮(pi)(pi)(pi)、肝、胰、肺泡上皮(pi)(pi)(pi)。 3. 游走型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(wandering cell type) 特(te)(te)點(dian)(dian):在(zai)(zai)(zai)支(zhi)持(chi)(chi)物(wu)上散在(zai)(zai)(zai)生(sheng)(sheng)(sheng)(sheng)長(chang),不(bu)(bu)連接成(cheng)(cheng)片,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)質伸出(chu)偽足或(huo)(huo)突起(qi)呈活躍的(de)(de)(de)(de)游走或(huo)(huo)變形(xing)(xing)(xing)(xing)運(yun)動,速度(du)快不(bu)(bu)規則(ze)(ze), 密度(du)大連接成(cheng)(cheng)片呈多角(jiao)形(xing)(xing)(xing)(xing)不(bu)(bu)易(yi)與(yu)其(qi)他類型(xing)的(de)(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao) 區別。 4. 多形(xing)(xing)(xing)(xing)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(polymorphic cell type) 特(te)(te)點(dian)(dian): 無(wu)規律(lv)的(de)(de)(de)(de)形(xing)(xing)(xing)(xing)態(tai),如神經(jing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)。 懸浮型(xing) 特(te)(te)點(dian)(dian): 不(bu)(bu)貼附(fu)(fu)在(zai)(zai)(zai)支(zhi)持(chi)(chi)物(wu)生(sheng)(sheng)(sheng)(sheng)長(chang),胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)體(ti)(ti)圓形(xing)(xing)(xing)(xing),在(zai)(zai)(zai)培(pei)(pei)養(yang)(yang)液中(zhong)生(sheng)(sheng)(sheng)(sheng)長(chang)空間大,可長(chang)時(shi)間的(de)(de)(de)(de)生(sheng)(sheng)(sheng)(sheng)長(chang),繁殖旺盛便(bian)于做(zuo)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)代謝研究。S180肉瘤、血液里(li)的(de)(de)(de)(de)白細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、K562、 HL-60。 分(fen)型(xing)的(de)(de)(de)(de)目的(de)(de)(de)(de): 方(fang)便(bian)描述細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)培(pei)(pei)養(yang)(yang)過程(cheng)中(zhong)的(de)(de)(de)(de)形(xing)(xing)(xing)(xing)態(tai)變化(hua)。培(pei)(pei)養(yang)(yang)條(tiao)件好時(shi),細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)相對穩定,可反(fan)映出(chu)起(qi)源、正常異常的(de)(de)(de)(de)區別,作為判定細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)生(sheng)(sheng)(sheng)(sheng)物(wu)學(xue)性狀(zhuang)指標。 強調:一(yi)般形(xing)(xing)(xing)(xing)態(tai)并不(bu)(bu)是(shi)一(yi)項可靠指標要(yao)受各方(fang)面(mian)影響。 如:反(fan)復(fu)開關溫箱、溫度(du)、C02的(de)(de)(de)(de)濃(nong)度(du)、培(pei)(pei)養(yang)(yang)基(ji)變堿、支(zhi)原體(ti)(ti)、pH值(zhi)。細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)接種時(shi)呈三(san)角(jiao)型(xing)狀(zhuang)態(tai),經(jing)培(pei)(pei)養(yang)(yang)一(yi)定時(shi)間后,細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)傳代前已形(xing)(xing)(xing)(xing)成(cheng)(cheng)上皮(pi)(pi)(pi)型(xing)細(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)。

KG-1a細胞(bao),人(ren)急性骨髓白血病細胞(bao)      25T

MV4-11細(xi)胞,人急性(xing)白血病細(xi)胞    25T

HL-60細(xi)胞,人急性早幼(you)粒白血病細(xi)胞    25T

U937細胞,人急性早幼粒白血(xue)病細胞     25T

MOLM-13細(xi)(xi)胞,人(ren)急性髓系白血病細(xi)(xi)胞    25T

MOLM-14細(xi)胞,人急性髓系白血(xue)病細(xi)胞    25T

K562細胞,人慢性髓系白血(xue)病細胞  25T

HL-60細胞,人(ren)急性早幼(you)粒白血病細胞    25T

CCRF-CEM細胞(bao),人急性淋巴細胞(bao)白血病T淋巴細胞(bao)  25T

HS 505.T細胞,慢性淋巴細胞白(bai)血(xue)病(bing)(CLL)細胞   25T

 

 

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