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供貨周期	現(xiàn)貨	規(guī)格	T25培養(yǎng)瓶x1 1.5ml凍存管x2
貨號(hào)	BFN608006464	應(yīng)用領(lǐng)域	醫(yī)療衛(wèi)生,生物產(chǎn)業(yè)
主要用途	僅供科研

細(xì)胞名稱	人自然殺傷性細(xì)胞NK92
貨物編碼	BFN608006464
產(chǎn)品規(guī)格	T25培養(yǎng)瓶x1	1.5ml凍存管x2
細(xì)胞數(shù)量	1x10^6	1x10^6
保存溫度	37℃	-198℃
運(yùn)輸方式	常溫保溫運(yùn)輸	干冰運(yùn)輸
安全等級(jí)	1
用途限制	僅供科研 3類

培養(yǎng)體系	The base medium for this cell line is Alpha Minimum Essential medium without ribonucleosides and deoxyribonucleosides but with 2 mM L-glutamine and 1.5 g/L sodium bicarbonate . To make the complete growth medium, add the following components to the base medium: 0.2 mM inositol; 0.1 mM 2-mercaptoethanol; 0.02 mM folic acid; 100-200 U/ml recombinant IL-2; adjust to a final concentration of 12.5% horse serum and 12.5% fetal bovine serum.
培養(yǎng)溫度	37℃	二氧化碳濃度	5%
簡(jiǎn)介	人自然殺傷性細(xì)胞NK92是從一位患有急進(jìn)性非霍奇金淋巴瘤的50歲白人男性外周血單核細(xì)胞衍生來(lái)的一株白細(xì)胞介素-2依賴型NK細(xì)胞株。NK-92MI是轉(zhuǎn)染得到的源自NK-92的IL-2非依賴的NK細(xì)胞株，親本細(xì)胞通過(guò)微粒體基因轉(zhuǎn)化法用逆轉(zhuǎn)錄病毒MFG-hIL-2載體攜帶的人IL-2cDNA進(jìn)行轉(zhuǎn)化。可能由于載體整合到基因組DNA中，轉(zhuǎn)化是穩(wěn)定的。這株細(xì)胞對(duì)很多惡性細(xì)胞有細(xì)胞毒性；鉻釋放試驗(yàn)顯示它能殺死K562和Daudi細(xì)胞。NK-92細(xì)胞有以下特征：CD2 +, CD7 +, CD11a +, CD28 + , CD45 +, CD54 +, CD56 +, CD1 -, CD3 -, CD4 -, CD5 -, CD8 -, CD10 -, CD14 -, CD16 -, CD19 -, CD20 -, CD23 -, CD34 -, HLA-DR -。另本庫(kù)保藏有NK92MI細(xì)胞。二者均引種自ATCC。
補(bǔ)充	NK92細(xì)胞培養(yǎng)時(shí)，會(huì)有零星聚團(tuán)現(xiàn)象，此時(shí)吹打散開即可。處于平臺(tái)期的細(xì)胞，會(huì)有少量呈現(xiàn)略不規(guī)則的圓形，此為正?，F(xiàn)象。血清建議使用BCK北美胎牛血清（貨號(hào)BCK041）與BCK馬血清（貨號(hào)BCK045)。
注釋	Part of: LL-100 blood cancer cell line panel. Characteristics: Laboratory use as a standard cell line for antibody-dependent cell-mediated cytotoxicity (ADCC) testing. Also bBeing developed for cellular adoptive immunotherapy for cancers and viral infections. Characteristics: IL2 dependent. Characteristics: Does not express FCGR3A/CD16. Doubling time: 35.6 +- 6.1 hours (PubMed=31126350); ~40-50 hours (DSMZ). Transformant: NCBI_TaxID; 10376; Epstein-Barr virus (EBV). Omics: Deep exome analysis. Omics: Deep RNAseq analysis. Omics: Transcriptome analysis. Miscellaneous: Neukoplast is used as a trademark to refer to NK-92 cells that are available for non-human research applications while aNK is used as a trademark to refer to cells from the cGMP-grade NK-92 cell-line that is in use for therapeutic human testing. Caution: This cell line is exclusively owned and controlled by NantKwest, Inc. NantKwest and its affiliate, Brink Biologics, Inc. are the sole authorized distributors for both commercial and non-commercial research requestors. There are no other authorized commercial and noncommercial suppliers of NK-92 cells. Contact NantKwest and Brink Biologics for information concerning current inventory and cell line use and support. Derived from sampling site: Peripheral blood.
STR信息	Amelogenin X,Y CSF1PO 11,12 D5S818 12,13 D7S820 10,11 D13S317 9,12 D16S539 11,12 TH01 6,9.3 TPOX 8 vWA 18 (ATCC) 16,18 (DSMZ)
參考文獻(xiàn)	PubMed=25586472; DOI=10.1038/ncomms7025 Kucuk C., Jiang B., Hu X., Zhang W., Chan J.K.C., Xiao W., Lack N., Alkan C., Williams J.C., Avery K.N., Kavak P., Scuto A., Sen E., Gaulard P., Staudt L.M., Iqbal J., Zhang W., Cornish A., Gong Q., Yang Q., Sun H., d'Amore F., Leppa S., Liu W., Fu K., de Leval L., McKeithan T., Chan W.C. Activating mutations of STAT5B and STAT3 in lymphomas derived from gammadelta-T or NK cells. Nat. Commun. 6:6025-6025(2015) PubMed=26559813; DOI=10.1007/s00262-015-1761-x Suck G., Odendahl M., Nowakowska P., Seidl C., Wels W.S., Klingemann H.-G., Tonn T. NK-92: an 'off-the-shelf therapeutic' for adoptive natural killer cell-based cancer immunotherapy. Cancer Immunol. Immunother. 65:485-492(2016) PubMed=31126350; DOI=10.1186/s40425-019-0612-2 Yang H.G., Kang M.C., Kim T.Y., Hwang I., Jin H.T., Sung Y.C., Eom K.-S., Kim S.W. Discovery of a novel natural killer cell line with distinct immunostimulatory and proliferative potential as an alternative platform for cancer immunotherapy. J. Immunother. Cancer 7:138-138(2019) PubMed=31160637; DOI=10.1038/s41598-019-44491-x Quentmeier H., Pommerenke C., Dirks W.G., Eberth S., Koeppel M., MacLeod R.A.F., Nagel S., Steube K., Uphoff C.C., Drexler H.G. The LL-100 panel: 100 cell lines for blood cancer studies. Sci. Rep. 9:8218-8218(2019)

驗(yàn)收細(xì)胞注意事項(xiàng)

1、收到人自然殺傷性細(xì)胞NK92細(xì)胞，請(qǐng)查看瓶子是否有破裂，培養(yǎng)基是否漏出，是否渾濁，如有請(qǐng)盡快聯(lián)系。

2、收到人自然殺傷性細(xì)胞NK92細(xì)胞，如包裝完好，請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞。,由于運(yùn)輸過(guò)程中的問(wèn)題，細(xì)胞培養(yǎng)瓶中的貼壁細(xì)胞有可能從瓶壁中脫落下來(lái)，顯微鏡下觀察會(huì)出現(xiàn)細(xì)胞懸浮的情況，出現(xiàn)此狀態(tài)時(shí)，請(qǐng)不要打開細(xì)胞培養(yǎng)瓶，應(yīng)立即將培養(yǎng)瓶置于細(xì)胞培養(yǎng)箱里靜止 3-5 小時(shí)左右，讓細(xì)胞先穩(wěn)定下，再于顯微鏡下觀察，此時(shí)多數(shù)細(xì)胞會(huì)重新貼附于瓶壁。如細(xì)胞仍不能貼壁，請(qǐng)用臺(tái)盼藍(lán)染色法鑒定細(xì)胞活力，如臺(tái)盼藍(lán)染色證實(shí)細(xì)胞活力正常請(qǐng)按懸浮細(xì)胞的方法處理。

3、收到人自然殺傷性細(xì)胞NK92細(xì)胞后，請(qǐng)鏡下觀察細(xì)胞，用恰當(dāng)方式處理細(xì)胞。若懸浮的細(xì)胞較多，請(qǐng)離心收集細(xì)胞，接種到一個(gè)新的培養(yǎng)瓶中。棄掉原液，使用新鮮配制的培養(yǎng)基，使用進(jìn)口胎牛血清。剛接到細(xì)胞，若細(xì)胞不多時(shí) 血清濃度可以加到 15％去培養(yǎng)。若細(xì)胞迏到 80%左右，血清濃度還是在 10％。

4、收到人自然殺傷性細(xì)胞NK92細(xì)胞時(shí)如無(wú)異常情況，請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞密度，如為貼壁細(xì)胞，未超過(guò)80%匯合度時(shí)，將培養(yǎng)瓶中培養(yǎng)基吸出，留下 5-10ML 培養(yǎng)基繼續(xù)培養(yǎng)：超過(guò) 80%匯合度時(shí)，請(qǐng)按細(xì)胞培養(yǎng)條件傳代培養(yǎng)。如為懸浮細(xì)胞，吸出培養(yǎng)液，1000 轉(zhuǎn)/分鐘離心 3 分鐘，吸出上清，管底細(xì)胞用新鮮培養(yǎng)基懸浮細(xì)胞后移回培養(yǎng)瓶。

5、將培養(yǎng)瓶置于 37℃培養(yǎng)箱中培養(yǎng)，蓋子微微擰松。吸出的培養(yǎng)基可以保存在滅菌過(guò)的瓶子里，存放于 4℃冰箱，以備不時(shí)之需。

6、24 小時(shí)后，人自然殺傷性細(xì)胞NK92細(xì)胞形態(tài)已恢復(fù)并貼滿瓶壁，即可傳代。（貼壁細(xì)胞）將培養(yǎng)瓶里的培養(yǎng)基倒去，加 3-5ml（以能覆蓋細(xì)胞生長(zhǎng)面為準(zhǔn)）PBS 或 Hanks’液洗滌后棄去。加 0.5-1ml 0.25%含 EDTA 的胰酶消化，消化時(shí)間以具體細(xì)胞為準(zhǔn)，一般 1-3 分鐘，不超過(guò) 5 分鐘?？梢苑?/span>入37℃培養(yǎng)箱消化。輕輕晃動(dòng)瓶壁，見細(xì)胞脫落下來(lái)，加入 3-5ml 培養(yǎng)基終止消化。用移液管輕輕吹打瓶壁上的細(xì)胞，使之*脫落，然后將溶液吸入離心管內(nèi)離心，1000rpm/5min。棄上清，視細(xì)胞數(shù)量決定分瓶數(shù)，一般一傳二，如細(xì)胞量多可一傳三，有些細(xì)胞不易傳得過(guò)稀，有些生長(zhǎng)較快的細(xì)胞則可以多傳幾瓶，以具體細(xì)胞和經(jīng)驗(yàn)為準(zhǔn)。（懸浮細(xì)胞）用移液管輕輕吹打瓶壁，直接將溶液吸入離心管離心即可。