NCI-H460大細(xi)胞肺(fei)癌細(xi)胞慧穎生(sheng)物一站(zhan)式(shi)采(cai)購商城，細(xi)胞株(zhu)細(xi)胞系，耐藥株(zhu)，STR鑒定，各(ge)品(pin)牌進口(kou)胎牛(niu)血清，培養(yang)基，胰酶(mei)，雙(shuang)抗(kang)，*培養(yang)基，PBS緩(huan)沖液，DMSO，細(xi)胞生(sheng)長因(yin)子等，種類齊全(quan)，*！

細(xi)(xi)胞(bao)(bao)培養(yang)技(ji)術(shu)(shu)屬于(yu)細(xi)(xi)胞(bao)(bao)工程范(fan)疇(chou)，細(xi)(xi)胞(bao)(bao)工程包(bao)括細(xi)(xi)胞(bao)(bao)融合技(ji)術(shu)(shu)、單克隆抗體(ti)的(de)制備(bei)、核移(yi)植技(ji)術(shu)(shu)（克隆）等,而這些技(ji)術(shu)(shu)中都有(you)細(xi)(xi)胞(bao)(bao)培養(yang)過(guo)程,所以說細(xi)(xi)胞(bao)(bao)培養(yang)是生物學研究(jiu)的(de)zui基本技(ji)術(shu)(shu)之一。

上海(hai)慧穎生物(wu)供應NCI-H460大細胞(bao)肺癌(ai)細胞(bao)，活性好、存活率高。隨著細胞生物(wu)學尤(you)其是干細胞研(yan)究(jiu)的(de)快速發展(zhan)，在(zai)細胞治療(liao)領域已經(jing)(jing)進(jin)行了(le)大量的(de)探(tan)索，取得(de)了(le)一(yi)系(xi)列的(de)令人振奮的(de)成(cheng)果，且部分(fen)基礎(chu)研(yan)究(jiu)成(cheng)果已經(jing)(jing)在(zai)心(xin)血管(guan)系(xi)統(tong)疾(ji)病 、神經(jing)(jing)系(xi)統(tong)疾(ji)病、肌肉骨骼相關(guan)疾(ji)病、糖尿病等。

養細(xi)胞經驗分享(xiang)：

1.細(xi)胞形態(tai)觀(guan)察。細(xi)胞膜(mo)(mo)(mo)是每天必須觀(guan)察的(de)，看是否清(qing)晰(xi)，細(xi)胞的(de)死亡往(wang)往(wang)是從細(xi)胞膜(mo)(mo)(mo)的(de)破(po)裂開始的(de)，早期的(de)表現就是細(xi)胞膜(mo)(mo)(mo)某一點出現欠完整或者毛發(fa)影。

2.細胞(bao)往往會有“抱團(tuan)”現象，抱團(tuan)生長(chang)或(huo)死亡(wang)，個人(ren)認(ren)為應該勤換液或(huo)傳代，盡量(liang)避 免(mian)細胞(bao)抱團(tuan)。

3.細(xi)(xi)胞(bao)生長迅速，強(qiang)烈建議用(yong)較大的培養(yang)瓶(ping)養(yang)，用(yong)小瓶(ping)養(yang)的話，換液不及時，很容易死 亡。細(xi)(xi)胞(bao)的生長速度一般為2天左右(you)就(jiu)需要傳代，并且一般為一傳三(san)，或一傳四(si)。

4.培(pei)養液：1640|DMEM|MEM|IMDM|a-MEM培(pei)養液，1L一般加碳酸(suan)氫(qing)鈉2g，并加雙抗，胎牛血清濃度*為12%，zui低不低于10%，有些細胞(bao)對胎牛血清高度依(yi)賴(lai)，比如WRO細胞(bao)，HT-22細胞(bao)，HA-VSMC細(xi)胞，NK-92細(xi)胞等。培養液PH應調為(wei)7.20-7.40之間。

5.防污染(ran)。L-02細(xi)胞生長(chang)迅速，較少(shao)發生污染(ran)，但常規(gui)措施應該做好。比如無菌操作，酒精(jing) 消毒等，培(pei)養瓶口建(jian)議(yi)過火，包括瓶蓋，應過火4秒左右。

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

7.復(fu)蘇。調節水浴(yu)箱溫度(du)為42℃，并提(ti)前在試管(guan)(guan)內放置(zhi)10倍培養液(ye)，使培養液(ye)溫度(du)與室溫一 致。1分鐘內融化細(xi)胞(bao)(禁止搖晃(huang)凍(dong)存管(guan)(guan))，移取(qu)至試管(guan)(guan)內，動(dong)作要輕(qing)，從試管(guan)(guan)管(guan)(guan)底(di)開始慢(man)(man)慢(man)(man)上提(ti) 移液(ye)管(guan)(guan)，使細(xi)胞(bao)在培養液(ye)內分布均勻，離心，洗(xi)2次。

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其(qi)它經驗概(gai)括：

細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)類型(xing)(xing)(xing) 編輯 細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)類型(xing)(xing)(xing)根據是(shi)否附(fu)于支持(chi)物(wu)上(shang)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)的(de)(de)(de)(de)(de)(de)特(te)(te)性，分(fen)為(wei) 貼附(fu)型(xing)(xing)(xing)、懸(xuan)浮(fu)型(xing)(xing)(xing)。 貼附(fu)型(xing)(xing)(xing) 細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)貼附(fu)在(zai)(zai)(zai)(zai)(zai)支持(chi)物(wu)表面(mian)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)，只依賴貼附(fu)才能生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)的(de)(de)(de)(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)叫做貼附(fu)型(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(Anchorrage-dependent cells)這種(zhong)(zhong)現象與(yu)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)分(fen)化(hua)有關。 貼附(fu)型(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)(de)(de)(de)分(fen)型(xing)(xing)(xing) 1.成(cheng)纖(xian)維(wei)型(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)胚(pei)層(ceng) 特(te)(te)點(dian)：與(yu)體(ti)內(nei)成(cheng)纖(xian)維(wei)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)形(xing)(xing)(xing)態(tai)相(xiang)似,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)體(ti)梭型(xing)(xing)(xing)或(huo)不規則三(san)角形(xing)(xing)(xing),中(zhong)(zhong)央有圓形(xing)(xing)(xing)核(he),胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)質向(xiang)外(wai)伸出(chu)2～3個長(chang)(chang)短(duan)不同的(de)(de)(de)(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)(zai)(zai)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)時(shi)(shi)呈(cheng)放射狀,漩渦或(huo)火(huo)焰(yan)狀走(zou)行。 起源(yuan)：細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)來自(zi)中(zhong)(zhong)胚(pei)層(ceng)間充質組織(zhi)。 除真正的(de)(de)(de)(de)(de)(de)成(cheng)纖(xian)維(wei)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、心(xin)肌、平(ping)滑(hua)肌、成(cheng)骨細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)、血(xue)管內(nei)皮(pi)(pi)(pi)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)(zai)(zai)培(pei)養(yang)(yang)時(shi)(shi)成(cheng)為(wei)成(cheng)纖(xian)維(wei)型(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)是(shi)一種(zhong)(zhong)慣稱,與(yu)體(ti)內(nei)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)不同。 2.上(shang)皮(pi)(pi)(pi)型(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(ceng)。 特(te)(te)點(dian)：扁(bian)平(ping)不規則多(duo)角型(xing)(xing)(xing)、圓形(xing)(xing)(xing)核(he)、細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)緊(jin)密相(xiang)連成(cheng)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)增(zeng)殖數(shu)目增(zeng)多(duo)時(shi)(shi),整(zheng)個上(shang)皮(pi)(pi)(pi)膜隨之移動。邊緣細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)很少(shao)脫離細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)群而單(dan)獨活(huo)動“ 拉網”現象與(yu)起源(yuan)內(nei)外(wai)胚(pei)層(ceng)組織(zhi)有關。 組織(zhi): 皮(pi)(pi)(pi)膚表皮(pi)(pi)(pi)及其衍生(sheng)(sheng)(sheng)(sheng)物(wu)(汗腺、皮(pi)(pi)(pi)脂腺)消化(hua)道分(fen)上(shang)皮(pi)(pi)(pi)、肝、胰、肺泡上(shang)皮(pi)(pi)(pi)。 3. 游(you)走(zou)型(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(te)(te)點(dian)：在(zai)(zai)(zai)(zai)(zai)支持(chi)物(wu)上(shang)散在(zai)(zai)(zai)(zai)(zai)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang),不連接成(cheng)片,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)質伸出(chu)偽足或(huo)突起呈(cheng)活(huo)躍的(de)(de)(de)(de)(de)(de)游(you)走(zou)或(huo)變(bian)形(xing)(xing)(xing)運動,速度快(kuai)不規則, 密度大(da)(da)連接成(cheng)片呈(cheng)多(duo)角形(xing)(xing)(xing)不易與(yu)其他類型(xing)(xing)(xing)的(de)(de)(de)(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao) 區(qu)別。 4. 多(duo)形(xing)(xing)(xing)型(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(te)(te)點(dian)： 無規律的(de)(de)(de)(de)(de)(de)形(xing)(xing)(xing)態(tai)，如(ru)(ru)神(shen)經(jing)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) 特(te)(te)點(dian)： 不貼附(fu)在(zai)(zai)(zai)(zai)(zai)支持(chi)物(wu)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)，胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)體(ti)圓形(xing)(xing)(xing)，在(zai)(zai)(zai)(zai)(zai)培(pei)養(yang)(yang)液(ye)中(zhong)(zhong)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)空間大(da)(da)，可長(chang)(chang)時(shi)(shi)間的(de)(de)(de)(de)(de)(de)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)，繁殖旺盛便于做細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)代(dai)謝研究。S180肉瘤、血(xue)液(ye)里的(de)(de)(de)(de)(de)(de)白(bai)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)的(de)(de)(de)(de)(de)(de)目的(de)(de)(de)(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)(zai)(zai)培(pei)養(yang)(yang)過程中(zhong)(zhong)的(de)(de)(de)(de)(de)(de)形(xing)(xing)(xing)態(tai)變(bian)化(hua)。培(pei)養(yang)(yang)條(tiao)件(jian)好時(shi)(shi)，細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)相(xiang)對穩(wen)定，可反映出(chu)起源(yuan)、正常(chang)異常(chang)的(de)(de)(de)(de)(de)(de)區(qu)別，作為(wei)判定細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)生(sheng)(sheng)(sheng)(sheng)物(wu)學性狀指標。 強調(diao)：一般形(xing)(xing)(xing)態(tai)并不是(shi)一項可靠指標要受各方面(mian)影響。 如(ru)(ru)：反復開關溫箱(xiang)、溫度、C02的(de)(de)(de)(de)(de)(de)濃度、培(pei)養(yang)(yang)基變(bian)堿(jian)、支原體(ti)、pH值。細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)(zai)接種(zhong)(zhong)時(shi)(shi)呈(cheng)三(san)角型(xing)(xing)(xing)狀態(tai)，經(jing)培(pei)養(yang)(yang)一定時(shi)(shi)間后，細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)(zai)傳(chuan)代(dai)前已形(xing)(xing)(xing)成(cheng)上(shang)皮(pi)(pi)(pi)型(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)(xi)胞(bao)，人急性骨(gu)髓白血病(bing)細(xi)(xi)胞(bao)      25T

MV4-11細胞，人(ren)急性白血病細胞    25T

HL-60細胞，人急(ji)性(xing)早幼(you)粒白血病細胞    25T

U937細(xi)胞，人急性早幼粒白(bai)血病(bing)細(xi)胞     25T

MOLM-13細(xi)胞，人(ren)急性髓系白(bai)血(xue)病細(xi)胞    25T

MOLM-14細胞，人急性髓系白(bai)血(xue)病細胞    25T

K562細(xi)胞(bao)，人(ren)慢性髓系白血病細(xi)胞(bao)  25T

HL-60細胞，人急(ji)性早幼粒白血病細胞    25T

CCRF-CEM細(xi)胞，人急(ji)性淋(lin)巴(ba)細(xi)胞白血病T淋(lin)巴(ba)細(xi)胞  25T

HS 505.T細胞，慢性淋巴(ba)細胞白(bai)血病（CLL）細胞   25T

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