Corning NK細(xì)胞活化擴增培養(yǎng)基套裝 NK細(xì)胞體外擴增試劑盒

Corning康寧公司NK細(xì)胞活化擴增培養(yǎng)基產(chǎn)品組成

1. NK細(xì)胞活化培養(yǎng)基

2. NK細(xì)胞活化添加劑

3. NK細(xì)胞擴增培養(yǎng)基

4. 抗體預(yù)包被T75細(xì)胞培養(yǎng)瓶

5. 細(xì)胞培養(yǎng)袋

中文名稱：康寧細(xì)胞活化擴增培養(yǎng)基套裝 NK細(xì)胞體外擴增試劑盒

英文名稱：Corning® NK Kit for Activation and Expansion of Human Natural Killer Cells

Corning康寧公司 NK細(xì)胞活化擴增培養(yǎng)基產(chǎn)品特色

1. 擴增能力強，在14天內(nèi)可達到100倍擴增

2. 效應(yīng)細(xì)胞比例高，可達80％

3. 操作方便

介紹

自然殺傷（NK）細(xì)胞是可識別和殺死腫瘤和病毒感染的關(guān)鍵免疫細(xì)胞。 NK細(xì)胞是當(dāng)今免疫-治療常用的細(xì)胞。但是NK細(xì)胞培養(yǎng)操作難度大，對培養(yǎng)條件、培養(yǎng)技術(shù)要求很高，難以獲得高純度高效能的活性細(xì)胞。過度免疫-治療所必需的離體NK細(xì)胞很難獲得。 已經(jīng)報道了幾種NK激活/擴增方法，其使用擴增介質(zhì)加各種細(xì)胞因子（例如IL-2，IL-7和IL-15）。 然而，這些需要研究人員的進一步優(yōu)化。

Corning康寧公司NK試劑盒包含預(yù)先涂覆的T-75培養(yǎng)瓶，50mL NK原代培養(yǎng)基，1.8mL NK Primary補充劑和1L NK擴增培養(yǎng)基。 NK細(xì)胞活化擴增試劑盒前三個組分是專為NK細(xì)胞激活設(shè)計的，而NK擴增培養(yǎng)基用于NK細(xì)胞的擴增。 NK初級培養(yǎng)基和NK擴增培養(yǎng)基都使用高品質(zhì)試劑和cGMP級原料制造。 NK細(xì)胞體外擴增試劑盒中存在于培養(yǎng)基中的*蛋白質(zhì)是血清白蛋白和重組人-胰島素的可注射水平。Corning NK細(xì)胞活化擴增培養(yǎng)基套裝*，現(xiàn)貨供應(yīng)。

Corning康寧公司 NK細(xì)胞活化/擴增培養(yǎng)基套裝由多個組分組成，為NK細(xì)胞培養(yǎng)提供簡便的操作方案。*，現(xiàn)貨供應(yīng)。并且使用Corning NK細(xì)胞活化擴增培養(yǎng)基套裝所獲得細(xì)胞數(shù)量產(chǎn)量高，活性高，具有很高的殺傷效應(yīng)。

關(guān)于使用該試劑盒激活和擴增的細(xì)胞的增殖率，免疫表型和CCK-8細(xì)胞毒性測試的信息，請參閱康寧應(yīng)用說明書康寧NK細(xì)胞體外擴增試劑盒（CLS-CG-AN-427）進行人類天然殺傷細(xì)胞擴增。

Corning康寧公司 NK細(xì)胞活化擴增培養(yǎng)基套裝

NK細(xì)胞體外擴增試劑盒

Related Products

Corning® NK Kit for Activation and Expansion of Human Natural Killer Cells Introduction

Natural killer (Corning NK) cells are crucial immune cells that can recognize and kill tumors and virus infection. It is difficult to obtain the large numbers of Corning NK cells ex vivo that are necessary for adoptive immu-notherapy. Several methods of Corning NK activation/expansion have been reported that use expansion medium plus various cytokines (e.g., IL-2, IL-7, and IL-15). However, these require further optimization by researchers.

The Corning NK Expansion kit provides a complete set of materials and a standard operating procedure to expand Corning NK cells with high expansion capacity, high viability, and high cytotoxicity. Researchers have no need to further optimize it, and the procedure has mini-mal consumables cost. In this application note, we provide com-petitive data, including cell yields, cell viability, and positive Corning NK ratio and cytotoxicity capability in vitro, that compares the Corning NK expansion kit with other commercially available expansion media.

健康人的PBMC細(xì)胞接種數(shù)量為3x107 ，根據(jù)廠家建議進行細(xì)胞培養(yǎng)。使用Corning培養(yǎng)基時，0-5天使用NK活化培養(yǎng)基 (Primary Medium)，6-14天使用NK 擴增培養(yǎng)基 (NK Expansion Medium )添加1000 IU/mL IL-2 進行培養(yǎng)。細(xì)胞培養(yǎng)至14天收獲，進行細(xì)胞計數(shù)和腫瘤殺傷能力檢測。腫瘤殺傷實驗使用的靶細(xì)胞為K562細(xì)胞。

實驗證明Corning NK細(xì)胞活化擴增培養(yǎng)基套裝 NK細(xì)胞體外擴增試劑盒效果更好。*，現(xiàn)貨供應(yīng)。NK細(xì)胞在Corning培養(yǎng)基中能夠得到更有效的擴增。Corning培養(yǎng)基中獲得的NK細(xì)胞具有更高的殺傷能力。

NK細(xì)胞激活和擴增

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在室溫（RT）下將抗凝血液以400xg離心10分鐘，然后將等離子體（頂層）轉(zhuǎn)移到新的管中。

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熱自動等離子體在56℃滅活30分鐘，然后以800xg離心20分鐘以除去沉淀物。上清血漿應(yīng)儲存在4℃，這將在隨后的14個培養(yǎng)日內(nèi)消耗。

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加入等體積的PBS（不含鈣和鎂的磷酸鹽緩沖鹽水，Corning Cat。編號21-040-CV），作為去除的自身血漿的替代物，以保持恒定體積并輕輕地重懸血細(xì)胞。

注意：在PBS中預(yù)先加入0.1％人血清白蛋白（HSA）有助于維持血細(xì)胞活力。

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Corning NK細(xì)胞活化擴增培養(yǎng)基套裝根據(jù)制造商的指示，使用淋巴細(xì)胞分離培養(yǎng)基（LSM，Corning目錄號25-072-Cl）從上述血液樣品中制備外周血單核細(xì)胞（PBMC）。

注意：使用新鮮收集的人體血液（收集后2小時內(nèi)），以獲得更好的表現(xiàn);在使用前不要使用超過24小時吸取的血液。

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用不含鈣和鎂的至少5倍PBS洗滌PBMC。離心機為500 xg

在室溫下10分鐘收集PBMC。

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使用含有1.8mL NK Primary補充劑和10％自動等離子體的NK Primary培養(yǎng)基將PBMC稀釋至1×10 6個細(xì)胞/ mL。

用于激活和擴展人類天然殺傷細(xì)胞的Corning康寧公司NK細(xì)胞體外擴增試劑盒協(xié)議

保修/免責(zé)聲明：除非另有規(guī)定，Corning NK細(xì)胞活化擴增培養(yǎng)基套裝所有產(chǎn)品僅供研究使用。*，現(xiàn)貨供應(yīng)。NK細(xì)胞無血清培養(yǎng)基不適用于診斷或治療程序。NK細(xì)胞無血清培養(yǎng)基不適用于人類。康寧生命科學(xué)不對Corning NK細(xì)胞活化擴增培養(yǎng)基套裝在臨床或診斷應(yīng)用中的性能提出任何索賠。

紅榮微再（上海）生物工程技術(shù)有限公司主營：干細(xì)胞、精準(zhǔn)醫(yī)療、細(xì)胞治療、器官再生 四大板塊的產(chǎn)品，以“傳遞科學(xué)價值，服務(wù)科學(xué)研究"為宗旨，經(jīng)過近兩年的努力推廣，紅榮微再（上海）生物工程技術(shù)有限公司在全中國代理的CellArtis干細(xì)胞產(chǎn)品已經(jīng)成功銷往：上海市、北京市、重慶市、天津市、廣東省的廣州市、深圳市、珠海市、江西省的南昌市、四川省的成都市、綿陽、浙江省的杭州市、寧波、溫州、江蘇省的南京市、蘇州、無錫、泰州、湖南省的長沙、陜西省的西安市、楊凌、甘肅省的蘭州、寧夏的銀川、新疆的烏魯木齊、廣西的南寧市、山東省的濟南、青島、東北三省 遼寧省的沈陽市、大連市、吉林省的長春市、黑龍江省的哈爾濱市等中國絕大部分省市和自治區(qū)。

歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司：紅榮微再（上海）生物工程技術(shù)有限公司  ：152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348

健康人的PBMC細(xì)胞根據(jù)廠家建議進行細(xì)胞培養(yǎng)。細(xì)胞培養(yǎng)至14天收獲，進行細(xì)胞計數(shù)和檢測。細(xì)胞使用CD3 FITC /CD56 PE進行染色，之后使用BDAccuriTM C6 flow cytometer進行檢測。實驗證明效果更好。*，現(xiàn)貨供應(yīng)。

Corning康寧公司NK細(xì)胞活化擴增培養(yǎng)基英文使用方法介紹

Corning® NK Kit for Activation and Expansion of Human Natural Killer Cells

Materials and Methods

Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donor whole blood using lymphocyte separation medium (Corning Cat. No. 25-072-CI) according to the manufacturer’s instructions. The wash buffer was PBS (Corning Cat. No. 21-040-CV), and the culture vessel was a gas-permeable cell culture media bag (Corning Cat. No. 88-610-20). Interleukin 2 (IL-2, Corning Cat. No. 354043) was used as an in vitro cytokine for Corning NK cell activation and expansion.

Corning NK Cell Activation and Expansion

On Day 0, the auto-plasma was inactivated by heating at 56°C for 30 min. and centrifuged at 800 xg for 20 min. to remove the precipitates. The supernatant was collected and stored at 4°C for further use.

The PBMCs were washed once with at least 5-fold PBS buffer, and the sample was centrifuged at 500 xg for 10 min. at room temperature. The density of PBMCs was adjusted to 1 x 106 cells/mL using Corning KBM NK Primary medium containing 1.8 mL Corning KBM NK Primary supplement and 10% auto-plasma.

The pre-coated T-75 flask was washed carefully twice with PBS just before use. Thirty milliliters of the suspension of PBMCs were seeded into the pre-coated T-75 flask and incubated at 37°C in a humidified atmosphere of 5% CO2 in air.

Note: Before Day 6 when the medium turned yellow and the cell density was above 2 x 106 cells/mL, fresh KBM Corning NK Primary medium plus 10% auto-plasma were added to the cell suspension to ensure that cell density stayed within the range of 2.5 x 105 to 2.0 x 106 cells/mL.

On Day 6, all cells were centrifuged and re-suspended with an appropriate volume of Corning KBM NK Expansion medium containing 1,000 IU/mL IL-2 and 10% auto-plasma and then transferred to a new T-225 flask or gas-permeable culture bag.

In the following days,Corning KBM NK Expansion medium containing 1,000 IU/mL IL-2 was added every 2 to 3 days based on the cell proliferation status to ensure that cell density stayed within 2.5 x 105 to 2.0 x 106 cells/mL.

The cells were harvested for CCK8 cytotoxic and immunophenotyping testing when the total cell number exceeded 2 x 109.

Note: The cells were gently dissociated, and the culture vessels were tapped softly to avoid cell damage and maintain cell viability throughout the entire experimental process.