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VSMC鼠胸主動脈平滑肌細胞,進口胎牛血清培養

發(fa)布時間(jian):2016-05-04瀏覽:972次

VSMC鼠胸主動(dong)脈平滑(hua)肌細胞,進口胎牛血清培養慧穎生(sheng)物(wu)一站(zhan)式(shi)采(cai)購商城,細(xi)胞(bao)株細(xi)胞(bao)系,耐(nai)藥(yao)株,STR鑒(jian)定(ding),各品牌(pai)進(jin)口(kou)胎牛血(xue)清(qing),培養(yang)基,胰(yi)酶,雙抗,*培養(yang)基,PBS緩(huan)沖(chong)液(ye),DMSO,細(xi)胞(bao)生(sheng)長因子等,種類齊全,*!

細(xi)胞培(pei)養(yang)技(ji)術(shu)屬于細(xi)胞工程(cheng)范疇,細(xi)胞工程(cheng)包(bao)括細(xi)胞融(rong)合技(ji)術(shu)、單克隆(long)(long)抗體的制備、核(he)移植(zhi)技(ji)術(shu)(克隆(long)(long))等,而這些技(ji)術(shu)中都(dou)有細(xi)胞培(pei)養(yang)過程(cheng),所(suo)以說(shuo)細(xi)胞培(pei)養(yang)是生物學研究(jiu)的zui基本技(ji)術(shu)之(zhi)一。

上海慧穎(ying)生(sheng)物供應VSMC鼠胸主動脈平(ping)滑肌細胞,進(jin)口胎(tai)牛血清(qing)培(pei)養,活性好、存(cun)活率高。隨著細(xi)胞生物(wu)學(xue)尤其是(shi)干細(xi)胞研(yan)究的(de)(de)快速(su)發展,在細(xi)胞治療領域(yu)已經(jing)進行了大量的(de)(de)探(tan)索,取得了一系(xi)列的(de)(de)令人(ren)振奮的(de)(de)成果,且部分基礎(chu)研(yan)究成果已經(jing)在心血管系(xi)統疾(ji)病(bing) 、神經(jing)系(xi)統疾(ji)病(bing)、肌肉骨骼(ge)相關疾(ji)病(bing)、糖(tang)尿病(bing)等。

養細胞經(jing)驗分(fen)享:

1.細(xi)胞(bao)形態觀察。細(xi)胞(bao)膜是每天必須觀察的,看是否(fou)清晰,細(xi)胞(bao)的死(si)亡往(wang)往(wang)是從細(xi)胞(bao)膜的破(po)裂開始的,早期(qi)的表(biao)現就是細(xi)胞(bao)膜某(mou)一(yi)點出現欠完整(zheng)或(huo)者毛發(fa)影。

2.細胞往往會有“抱(bao)團(tuan)”現(xian)象(xiang),抱(bao)團(tuan)生長或死亡(wang),個人認(ren)為(wei)應(ying)該勤換(huan)液或傳代(dai),盡(jin)量避(bi) 免細胞抱(bao)團(tuan)。

3.細(xi)胞生長迅速(su)(su),強烈(lie)建(jian)議用較大的培養(yang)瓶養(yang),用小瓶養(yang)的話,換(huan)液不(bu)及(ji)時,很(hen)容易死 亡。細(xi)胞的生長速(su)(su)度一(yi)般為(wei)(wei)2天左右就需要(yao)傳(chuan)代(dai),并且一(yi)般為(wei)(wei)一(yi)傳(chuan)三,或一(yi)傳(chuan)四。

4.培養(yang)液:1640|DMEM|MEM|IMDM|a-MEM培養(yang)液,1L一般加(jia)碳酸氫鈉(na)2g,并加(jia)雙抗,胎(tai)牛血清(qing)濃度*為12%,zui低不低于10%,有(you)些(xie)細胞(bao)(bao)對(dui)胎(tai)牛血清(qing)高度依賴(lai),比如WRO細胞(bao)(bao),HT-22細胞(bao)(bao),HA-VSMC細胞(bao),NK-92細胞(bao)等。培(pei)養液PH應調為7.20-7.40之間。

5.防污(wu)染(ran)。L-02細胞生長迅速,較少發生污(wu)染(ran),但常規措施應該(gai)做好。比(bi)如無菌操作(zuo),酒精 消毒等,培養(yang)瓶口建議過火(huo),包括瓶蓋,應過火(huo)4秒左右。

6.凍存(cun)(cun)。-20一小時,-80次夜,液氮(dan)長(chang)(chang)期(qi)保存(cun)(cun)。強烈建(jian)議液氮(dan)長(chang)(chang)期(qi)保存(cun)(cun),盡(jin)量(liang)不要在-80長(chang)(chang)期(qi) 保存(cun)(cun),死(si)亡率很高(gao)。

7.復蘇。調(diao)節水浴箱溫(wen)度(du)為42℃,并提(ti)前在試(shi)管(guan)內(nei)放置10倍培養(yang)液,使(shi)培養(yang)液溫(wen)度(du)與室溫(wen)一(yi) 致。1分鐘(zhong)內(nei)融化(hua)細胞(禁止搖晃凍存管(guan)),移(yi)取至試(shi)管(guan)內(nei),動作要輕,從試(shi)管(guan)管(guan)底開始慢慢上提(ti) 移(yi)液管(guan),使(shi)細胞在培養(yang)液內(nei)分布(bu)均勻,離(li)心,洗2次。

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其它(ta)經驗概(gai)括:

細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)類(lei)型(xing)(xing) 編輯 細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)類(lei)型(xing)(xing)根據是(shi)(shi)否附(fu)于支(zhi)持(chi)(chi)物(wu)(wu)(wu)上(shang)(shang)生(sheng)(sheng)長(chang)(chang)的(de)(de)(de)特(te)性,分(fen)為 貼(tie)附(fu)型(xing)(xing)、懸浮型(xing)(xing)。 貼(tie)附(fu)型(xing)(xing) 細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)貼(tie)附(fu)在(zai)(zai)(zai)(zai)支(zhi)持(chi)(chi)物(wu)(wu)(wu)表面生(sheng)(sheng)長(chang)(chang),只依賴貼(tie)附(fu)才(cai)能生(sheng)(sheng)長(chang)(chang)的(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)叫做(zuo)貼(tie)附(fu)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(Anchorrage-dependent cells)這種(zhong)現(xian)象(xiang)與(yu)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)分(fen)化(hua)有(you)關。 貼(tie)附(fu)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)的(de)(de)(de)分(fen)型(xing)(xing) 1.成(cheng)纖(xian)(xian)維(wei)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao):(fibroblast) 來自中(zhong)(zhong)胚(pei)層(ceng) 特(te)點:與(yu)體(ti)內成(cheng)纖(xian)(xian)維(wei)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)形(xing)(xing)(xing)態(tai)相似,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)體(ti)梭型(xing)(xing)或(huo)(huo)不規(gui)(gui)則(ze)三角(jiao)(jiao)形(xing)(xing)(xing),中(zhong)(zhong)央有(you)圓(yuan)形(xing)(xing)(xing)核,胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)質(zhi)向(xiang)外(wai)伸出2~3個(ge)長(chang)(chang)短不同(tong)的(de)(de)(de)突起(qi)(qi)。細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)生(sheng)(sheng)長(chang)(chang)時呈(cheng)放射狀,漩渦(wo)或(huo)(huo)火焰(yan)狀走(zou)行。 起(qi)(qi)源(yuan):細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)來自中(zhong)(zhong)胚(pei)層(ceng)間(jian)充質(zhi)組(zu)(zu)織(zhi)。 除真正(zheng)的(de)(de)(de)成(cheng)纖(xian)(xian)維(wei)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)、心肌(ji)、平滑肌(ji)、成(cheng)骨細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)、血管內皮(pi)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)。 注意(yi):細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)培(pei)(pei)養時成(cheng)為成(cheng)纖(xian)(xian)維(wei)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)是(shi)(shi)一種(zhong)慣稱,與(yu)體(ti)內細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)不同(tong)。 2.上(shang)(shang)皮(pi)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(epithlium cell type) 來自外(wai)胚(pei)層(ceng)。 特(te)點:扁平不規(gui)(gui)則(ze)多(duo)角(jiao)(jiao)型(xing)(xing)、圓(yuan)形(xing)(xing)(xing)核、細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)緊密相連成(cheng)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)增殖(zhi)數目增多(duo)時,整個(ge)上(shang)(shang)皮(pi)膜隨之移動。邊緣細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)很(hen)少脫離細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)群而單(dan)獨活動“ 拉(la)網”現(xian)象(xiang)與(yu)起(qi)(qi)源(yuan)內外(wai)胚(pei)層(ceng)組(zu)(zu)織(zhi)有(you)關。 組(zu)(zu)織(zhi): 皮(pi)膚表皮(pi)及其(qi)衍生(sheng)(sheng)物(wu)(wu)(wu)(汗腺、皮(pi)脂腺)消化(hua)道分(fen)上(shang)(shang)皮(pi)、肝、胰、肺泡上(shang)(shang)皮(pi)。 3. 游(you)(you)走(zou)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(wandering cell type) 特(te)點:在(zai)(zai)(zai)(zai)支(zhi)持(chi)(chi)物(wu)(wu)(wu)上(shang)(shang)散在(zai)(zai)(zai)(zai)生(sheng)(sheng)長(chang)(chang),不連接成(cheng)片(pian)(pian),胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)質(zhi)伸出偽足或(huo)(huo)突起(qi)(qi)呈(cheng)活躍的(de)(de)(de)游(you)(you)走(zou)或(huo)(huo)變形(xing)(xing)(xing)運動,速度快不規(gui)(gui)則(ze), 密度大連接成(cheng)片(pian)(pian)呈(cheng)多(duo)角(jiao)(jiao)形(xing)(xing)(xing)不易(yi)與(yu)其(qi)他類(lei)型(xing)(xing)的(de)(de)(de)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao) 區別。 4. 多(duo)形(xing)(xing)(xing)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)(polymorphic cell type) 特(te)點: 無(wu)規(gui)(gui)律的(de)(de)(de)形(xing)(xing)(xing)態(tai),如神經細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)。 懸浮型(xing)(xing) 特(te)點: 不貼(tie)附(fu)在(zai)(zai)(zai)(zai)支(zhi)持(chi)(chi)物(wu)(wu)(wu)生(sheng)(sheng)長(chang)(chang),胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)體(ti)圓(yuan)形(xing)(xing)(xing),在(zai)(zai)(zai)(zai)培(pei)(pei)養液(ye)中(zhong)(zhong)生(sheng)(sheng)長(chang)(chang)空間(jian)大,可(ke)長(chang)(chang)時間(jian)的(de)(de)(de)生(sheng)(sheng)長(chang)(chang),繁殖(zhi)旺(wang)盛便于做(zuo)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)代謝研究。S180肉(rou)瘤、血液(ye)里(li)的(de)(de)(de)白細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)、K562、 HL-60。 分(fen)型(xing)(xing)的(de)(de)(de)目的(de)(de)(de): 方便描述細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)培(pei)(pei)養過程中(zhong)(zhong)的(de)(de)(de)形(xing)(xing)(xing)態(tai)變化(hua)。培(pei)(pei)養條件好(hao)時,細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)相對穩定,可(ke)反(fan)映出起(qi)(qi)源(yuan)、正(zheng)常異常的(de)(de)(de)區別,作為判(pan)定細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)生(sheng)(sheng)物(wu)(wu)(wu)學性狀指(zhi)標(biao)。 強調:一般形(xing)(xing)(xing)態(tai)并不是(shi)(shi)一項可(ke)靠指(zhi)標(biao)要受各方面影響(xiang)。 如:反(fan)復開關溫箱、溫度、C02的(de)(de)(de)濃度、培(pei)(pei)養基變堿、支(zhi)原體(ti)、pH值(zhi)。細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)接種(zhong)時呈(cheng)三角(jiao)(jiao)型(xing)(xing)狀態(tai),經培(pei)(pei)養一定時間(jian)后,細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)在(zai)(zai)(zai)(zai)傳代前已(yi)形(xing)(xing)(xing)成(cheng)上(shang)(shang)皮(pi)型(xing)(xing)細(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)(bao)(bao)(bao)。

KG-1a細(xi)胞,人急性骨髓白血病細(xi)胞      25T

MV4-11細(xi)胞,人急(ji)性白血病細(xi)胞    25T

HL-60細胞,人(ren)急性早幼粒白血病細胞    25T

U937細胞(bao),人急性(xing)早幼粒(li)白血病(bing)細胞(bao)     25T

MOLM-13細胞,人急性髓(sui)系白血病細胞    25T

MOLM-14細胞(bao),人急(ji)性髓(sui)系白血(xue)病細胞(bao)    25T

K562細胞(bao),人慢性髓系(xi)白(bai)血病細胞(bao)  25T

HL-60細(xi)胞,人(ren)急性早幼粒白血病(bing)細(xi)胞    25T

CCRF-CEM細(xi)胞(bao),人急性淋巴(ba)細(xi)胞(bao)白血(xue)病T淋巴(ba)細(xi)胞(bao)  25T

HS 505.T細胞(bao)(bao),慢性(xing)淋(lin)巴(ba)細胞(bao)(bao)白血(xue)病(CLL)細胞(bao)(bao)   25T

 

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