293T SV40永生化的人胚腎上皮細胞(bao)，進口胎牛血清(qing)培(pei)養(yang)慧(hui)穎生物一站式采購商城(cheng)，細胞(bao)株細胞(bao)系，耐藥株，STR鑒定，各品牌進口胎牛血清(qing)，培(pei)(pei)養(yang)基，胰酶，雙(shuang)抗(kang)，*培(pei)(pei)養(yang)基，PBS緩沖液，DMSO，細胞(bao)生長因子(zi)等，種類齊(qi)全，*！

細(xi)(xi)胞(bao)培(pei)養(yang)技(ji)術(shu)屬于(yu)細(xi)(xi)胞(bao)工(gong)程范(fan)疇(chou)，細(xi)(xi)胞(bao)工(gong)程包(bao)括(kuo)細(xi)(xi)胞(bao)融(rong)合技(ji)術(shu)、單(dan)克隆抗體的(de)制備、核移植技(ji)術(shu)（克隆）等(deng),而這些技(ji)術(shu)中都有細(xi)(xi)胞(bao)培(pei)養(yang)過(guo)程,所(suo)以說細(xi)(xi)胞(bao)培(pei)養(yang)是生物學研究的(de)zui基本(ben)技(ji)術(shu)之(zhi)一。

上(shang)海慧穎生物供應293T SV40永生化的人胚腎上皮細胞，進口(kou)胎牛(niu)血清培(pei)養，活性好、存(cun)活率高(gao)。隨著細胞(bao)(bao)生物學(xue)尤(you)其是干細胞(bao)(bao)研究的(de)快速發(fa)展，在細胞(bao)(bao)治療領域已(yi)經進行了大量的(de)探(tan)索，取得(de)了一系列的(de)令人振奮的(de)成果，且(qie)部分基礎研究成果已(yi)經在心血管(guan)系統疾病 、神經系統疾病、肌(ji)肉骨骼相關疾病、糖尿病等。

養細胞經驗分享：

1.細胞(bao)(bao)形態觀(guan)察(cha)。細胞(bao)(bao)膜是(shi)(shi)每天必須(xu)觀(guan)察(cha)的，看是(shi)(shi)否清晰，細胞(bao)(bao)的死(si)亡往往是(shi)(shi)從細胞(bao)(bao)膜的破裂(lie)開始(shi)的，早期(qi)的表現就是(shi)(shi)細胞(bao)(bao)膜某一點出現欠完整或者毛發影(ying)。

2.細胞往往會有“抱(bao)(bao)(bao)團(tuan)”現象，抱(bao)(bao)(bao)團(tuan)生(sheng)長或(huo)死亡，個人認為應該(gai)勤(qin)換液或(huo)傳代，盡(jin)量避(bi) 免(mian)細胞抱(bao)(bao)(bao)團(tuan)。

3.細(xi)胞生長迅速(su)，強烈建議用較大的(de)培養(yang)瓶養(yang)，用小瓶養(yang)的(de)話，換液(ye)不及時，很容易死 亡。細(xi)胞的(de)生長速(su)度一般為(wei)2天左右就(jiu)需要(yao)傳代，并且一般為(wei)一傳三(san)，或一傳四(si)。

4.培養(yang)液(ye)：1640|DMEM|MEM|IMDM|a-MEM培養(yang)液(ye)，1L一般加碳酸氫鈉2g，并加雙(shuang)抗，胎牛(niu)血清(qing)濃度(du)*為12%，zui低不低于10%，有些(xie)細(xi)胞對胎牛(niu)血清(qing)高度(du)依賴，比(bi)如WRO細(xi)胞，HT-22細(xi)胞，HA-VSMC細胞，NK-92細胞等(deng)。培養液PH應(ying)調為7.20-7.40之間。

5.防污(wu)染(ran)(ran)。L-02細(xi)胞生長(chang)迅速，較少發生污(wu)染(ran)(ran)，但常規措施應該做(zuo)好。比(bi)如無菌操作，酒(jiu)精 消(xiao)毒等，培養瓶口建議過(guo)火，包括瓶蓋，應過(guo)火4秒左右。

6.凍(dong)存。-20一小時(shi)，-80次(ci)夜(ye)，液(ye)氮(dan)長(chang)(chang)期(qi)保(bao)存。強烈(lie)建議液(ye)氮(dan)長(chang)(chang)期(qi)保(bao)存，盡量不要在-80長(chang)(chang)期(qi) 保(bao)存，死亡率(lv)很(hen)高(gao)。

7.復蘇。調節水(shui)浴箱溫度為42℃，并提前(qian)在試管(guan)內放(fang)置10倍培養液(ye)(ye)，使培養液(ye)(ye)溫度與室溫一 致。1分鐘(zhong)內融化細(xi)(xi)胞(禁止搖(yao)晃(huang)凍存管(guan))，移(yi)取至(zhi)試管(guan)內，動作要輕，從試管(guan)管(guan)底開始慢慢上提 移(yi)液(ye)(ye)管(guan)，使細(xi)(xi)胞在培養液(ye)(ye)內分布(bu)均勻，離心(xin)，洗(xi)2次。

相關(guan)熱賣產品(pin)：

其它經驗概括：

細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)類(lei)型(xing)(xing)(xing)(xing)(xing)(xing) 編(bian)輯 細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)類(lei)型(xing)(xing)(xing)(xing)(xing)(xing)根據是否附(fu)于支持物(wu)上(shang)生(sheng)長(chang)(chang)的特(te)性，分(fen)為 貼(tie)(tie)附(fu)型(xing)(xing)(xing)(xing)(xing)(xing)、懸浮型(xing)(xing)(xing)(xing)(xing)(xing)。 貼(tie)(tie)附(fu)型(xing)(xing)(xing)(xing)(xing)(xing) 細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)貼(tie)(tie)附(fu)在(zai)(zai)(zai)支持物(wu)表(biao)面生(sheng)長(chang)(chang)，只依(yi)賴(lai)貼(tie)(tie)附(fu)才能生(sheng)長(chang)(chang)的細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)叫做(zuo)貼(tie)(tie)附(fu)型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(Anchorrage-dependent cells)這(zhe)種現象與細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)分(fen)化有(you)(you)關(guan)。 貼(tie)(tie)附(fu)型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)的分(fen)型(xing)(xing)(xing)(xing)(xing)(xing) 1.成(cheng)纖維(wei)(wei)型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)：(fibroblast) 來自中胚(pei)層(ceng)(ceng) 特(te)點：與體(ti)內成(cheng)纖維(wei)(wei)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)形(xing)(xing)態相(xiang)似,胞(bao)(bao)(bao)(bao)(bao)(bao)體(ti)梭型(xing)(xing)(xing)(xing)(xing)(xing)或(huo)不(bu)(bu)(bu)(bu)(bu)規(gui)則三(san)角(jiao)形(xing)(xing),中央有(you)(you)圓形(xing)(xing)核,胞(bao)(bao)(bao)(bao)(bao)(bao)質向外(wai)(wai)伸(shen)(shen)出(chu)2～3個(ge)(ge)長(chang)(chang)短不(bu)(bu)(bu)(bu)(bu)同的突起(qi)(qi)。細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)生(sheng)長(chang)(chang)時(shi)呈放射狀,漩渦或(huo)火焰狀走(zou)(zou)行。 起(qi)(qi)源(yuan)：細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)來自中胚(pei)層(ceng)(ceng)間(jian)(jian)充(chong)質組(zu)織(zhi)。 除真正的成(cheng)纖維(wei)(wei)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)、心肌、平(ping)滑肌、成(cheng)骨細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)、血管內皮細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)。 注意：細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)培(pei)養時(shi)成(cheng)為成(cheng)纖維(wei)(wei)型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)是一種慣稱,與體(ti)內細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)不(bu)(bu)(bu)(bu)(bu)同。 2.上(shang)皮型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(epithlium cell type) 來自外(wai)(wai)胚(pei)層(ceng)(ceng)。 特(te)點：扁平(ping)不(bu)(bu)(bu)(bu)(bu)規(gui)則多(duo)(duo)角(jiao)型(xing)(xing)(xing)(xing)(xing)(xing)、圓形(xing)(xing)核、細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)緊(jin)密相(xiang)連(lian)成(cheng)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)增殖(zhi)(zhi)數目增多(duo)(duo)時(shi),整個(ge)(ge)上(shang)皮膜(mo)隨之移動(dong)。邊緣細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)很少脫離細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)群而單(dan)獨(du)活動(dong)“ 拉網(wang)”現象與起(qi)(qi)源(yuan)內外(wai)(wai)胚(pei)層(ceng)(ceng)組(zu)織(zhi)有(you)(you)關(guan)。 組(zu)織(zhi): 皮膚表(biao)皮及其衍生(sheng)物(wu)(汗腺、皮脂腺)消化道分(fen)上(shang)皮、肝、胰、肺泡(pao)上(shang)皮。 3. 游走(zou)(zou)型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(wandering cell type) 特(te)點：在(zai)(zai)(zai)支持物(wu)上(shang)散在(zai)(zai)(zai)生(sheng)長(chang)(chang),不(bu)(bu)(bu)(bu)(bu)連(lian)接(jie)成(cheng)片,胞(bao)(bao)(bao)(bao)(bao)(bao)質伸(shen)(shen)出(chu)偽足(zu)或(huo)突起(qi)(qi)呈活躍的游走(zou)(zou)或(huo)變(bian)形(xing)(xing)運動(dong),速度快不(bu)(bu)(bu)(bu)(bu)規(gui)則, 密度大(da)連(lian)接(jie)成(cheng)片呈多(duo)(duo)角(jiao)形(xing)(xing)不(bu)(bu)(bu)(bu)(bu)易與其他類(lei)型(xing)(xing)(xing)(xing)(xing)(xing)的細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao) 區別。 4. 多(duo)(duo)形(xing)(xing)型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(polymorphic cell type) 特(te)點： 無規(gui)律的形(xing)(xing)態，如(ru)神(shen)經(jing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)。 懸浮型(xing)(xing)(xing)(xing)(xing)(xing) 特(te)點： 不(bu)(bu)(bu)(bu)(bu)貼(tie)(tie)附(fu)在(zai)(zai)(zai)支持物(wu)生(sheng)長(chang)(chang)，胞(bao)(bao)(bao)(bao)(bao)(bao)體(ti)圓形(xing)(xing)，在(zai)(zai)(zai)培(pei)養液中生(sheng)長(chang)(chang)空間(jian)(jian)大(da)，可長(chang)(chang)時(shi)間(jian)(jian)的生(sheng)長(chang)(chang)，繁殖(zhi)(zhi)旺(wang)盛便(bian)(bian)于做(zuo)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)代(dai)謝研究。S180肉瘤、血液里(li)的白細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)、K562、 HL－60。 分(fen)型(xing)(xing)(xing)(xing)(xing)(xing)的目的： 方便(bian)(bian)描述(shu)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)培(pei)養過程中的形(xing)(xing)態變(bian)化。培(pei)養條件好時(shi)，細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)相(xiang)對穩定(ding)，可反(fan)映出(chu)起(qi)(qi)源(yuan)、正常異常的區別，作為判定(ding)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)生(sheng)物(wu)學(xue)性狀指標(biao)。 強(qiang)調：一般形(xing)(xing)態并(bing)不(bu)(bu)(bu)(bu)(bu)是一項可靠指標(biao)要受各方面影(ying)響。 如(ru)：反(fan)復(fu)開(kai)關(guan)溫箱、溫度、C02的濃(nong)度、培(pei)養基(ji)變(bian)堿、支原體(ti)、pH值。細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)接(jie)種時(shi)呈三(san)角(jiao)型(xing)(xing)(xing)(xing)(xing)(xing)狀態，經(jing)培(pei)養一定(ding)時(shi)間(jian)(jian)后，細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)傳代(dai)前已形(xing)(xing)成(cheng)上(shang)皮型(xing)(xing)(xing)(xing)(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)。

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