亚州一区二区三区中文字幕国产精品-日韩人妻一区二区三区蜜桃视频-亚洲中文字幕久久无码精品-午夜精品亚洲一区二区三区嫩草-日韩人妻一区二区三区蜜桃视频-国产一区二区精品高清在线观看-国产欧美日韩综合精品一区二区

武漢原生原代生物醫藥科技有限公司

PriCells: Identification and expansion of the tumorigenic lung cancer stem cell population

時間:2021-11-10 閱讀:236
分享:

PriCells: Identification and expansion of the tumorigenic lung cancer stem cell population    

Lung cancer contains a rare population of CD133+ cancer stem-like cells able to self-renew and generates an unlimited progeny of non-tumorigenic cells. The current technology allows the establishment of long-term lung cancer stem cell cultures from about one-third of the tumors.
1. Tumor samples were obtained.
2. Surgical specimens were washed several times and left overnight in culture system.
DMEM–F12 medium supplemented
high doses of penicillin/streptomycin
amphotericin B
3. Tissue dissociation was carried out by enzymatic digestion (20?μg/ml collagenase II) for 2?h at 37°C.
4. Recovered cells were cultured at clonal density in serum-free medium.
50?μg/ml insulin
100?μg/ml apo-transferrin
10?μg/ml putrescine
0.03?mM sodium selenite
2?μM progesterone
0.6% glucose
5?mM HEPES
0.1% sodium bicarbonate
0.4% BSA
glutamine and antibiotics
20?μg/ml EGF
10?μg/ml bFGF.
5. Flasks non-treated for tissue culture were used to reduce cell adherence and support growth as undifferentiated tumor spheres.
6. The medium was replaced or supplemented with fresh growth factors twice a week until cells started to grow forming floating aggregates.
7. Cultures were expanded by mechanical dissociation of spheres, followed by re-plating of both single cells and residual small aggregates in complete fresh medium.
8. Stem cell medium was replaced with Bronchial Epithelial Cell Growth Medium in tissue culture-treated flasks.
9. Identification: Antibodies used were PE-conjugated anti-CD133/1, PE-conjugated anti-CD133/2 or APC-conjugated anti-CD133/1, anti-CD56/N-CAM, FITC-conjugated anti-epithelial membrane antigen, anti-human CKs, anti-CEA, FITC-conjugated anti-CD34 and anti-CD45, anti-CD31.
10. Single-cell suspensions from lung cancer specimens were prepared.
11. After thawing, cells were double stained with PE-conjugated anti-CD133/1 antibody and FITC-conjugated anti-Ep-CAM antibody.
12. Purity of the CD133+ and CD133? cell populations was evaluated.

會員登錄

×

請輸入賬號

請輸入密碼

=

請輸驗證碼

收藏該商鋪

X
該信息已收藏!
標簽:
保存成功

(空格分隔,最多3個,單個標簽最多10個字符)

常用:

提示

X
您的留言已提交成功!我們將在第一時間回復您~
撥打電話 產品分類
在線留言
主站蜘蛛池模板: 杨浦区| 兴义市| 崇礼县| 福泉市| 泾川县| 平昌县| 大余县| 扶余县| 松潘县| 通渭县| 扶余县| 花莲市| SHOW| 山阳县| 江安县| 沁水县| 阜康市| 池州市| 象州县| 宁武县| 图木舒克市| 杂多县| 环江| 大港区| 上栗县| 葫芦岛市| 福清市| 梅州市| 随州市| 阿巴嘎旗| 沧州市| 雅江县| 枣庄市| 绿春县| 西安市| 望奎县| 大石桥市| 松桃| 左权县| 阿巴嘎旗| 灵山县|