久久久久亚洲精品男人的天堂,日本韩国男男作爱GAYWWW,特黄AAAAAAA片免费视频,av天堂电影网

武漢原生原代生物醫(yī)藥科技有限公司

PriCells: Isolation and transcription profiling of purified uncultured human stromal stem cells

時(shí)間:2022-1-18 閱讀:276
分享:

PriCells: Isolation and transcription profiling of purified uncultured human stromal stem cells      


Isolation of Stromal Vascular Cells from Human Adipose Tissue
1. Adipose tissue was obtained by liposuction from abdominal, hip, and thigh regions of healthy female donors aged 18–39.
2. The stromal vascular fraction (SVF) was separated from adipose tissue. Lipoaspirate (300–400 ml) was washed with Hanks' balanced salt solution containing antibiotics (100 IU/ml penicillin and 100 IU/ml streptomycin, Life Technologies-BRL) and 2.5 μg/ml amphotericin B.
3. Washed adipose tissue was digested for 2 h on a shaker at 37℃ in HBSS containing 0.2% collagenase.
4. Floating adipocytes were aspirated from pelleted SVF cells after centrifugation at 400 × g for 10 min.
5. Pellets were resuspended in red blood cell lysis buffer (2.06 g/l Tris base, 7.49 g/l NH4Cl, pH 7.2) for 10 min at room temperature.
6. After resuspending SVF cells in HBSS containing 2% fetal bovine serum (FBS), tissue clumps were allowed to settle for 1 min.
7. Suspended cells were passed through 100-μm and then 40-μm cell sieves. Cell suspensions (15 ml) were applied to Histopaque-1077 gradients (15 ml) in 50-ml tubes.
8. After centrifugation (400 × g, 30 min), cells at the gradient interface were collected, washed in HBSS, and passed through a 30-μm mesh. Cell counts and viability assessment were performed.

Isolation of ADASC from SVF
1. CD45+ cells were removed using magnetic beads coupled to mouse antihuman CD45 MAb and a superMACS magnet according to instructions provided by the manufacturer.
2. Magnetic beads were also used to split the remaining CD45- cells into CD31+ and CD31- cells.
3. CD45- cells were incubated with FITC-conjugated mouse anti-human CD31 MAb at 1:40 dilution for 15 min at 4℃. After washing, antibody-coated cells were incubated with anti-FITC Microbeads for 15 min at 4℃.
4. Microbead-coated CD31+ cells were retained in LS columns, allowing the initial isolation of CD31- cells, and later flushed out separately according to the manufacturer's protocol.
5. Flow cytometry was used to confirm separation. We never observed more than 8% contaminating cells in either population.

Culture of Freshly Isolated ADASC
1. Immediately after separation, CD31+ and CD31- cell subsets were washed and resuspended in DMEM/F12 containing 20% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 2.5 μg/ml amphotericin B.
2. Cells were added to 25-cm2 culture flasks (105 cells/flask) and cultured at 37℃ in an atmosphere of 5% CO2 in humid air.
3. After 7 d, attached cells were passaged by trypsinization and cultured in DMEM/F12 containing 10% FBS without amphotericin B. Medium was replaced every third day.
4. Generation of clonal cell lines from CD31- cells was achieved by culturing single fresh cells in separate wells.
5. Using a micropipette, a single cell was placed into each well of 48-well plates containing DMEM/F12 with 20% FBS, antibiotics, and 2.5 μg/ml amphotericin B.
6. Colony-forming ability was assessed after 21 d. At that time, colonies were passaged and cultured further in DMEM/F12 containing 10% FBS without amphotericin B.

會(huì)員登錄

×

請(qǐng)輸入賬號(hào)

請(qǐng)輸入密碼

=

請(qǐng)輸驗(yàn)證碼

收藏該商鋪

X
該信息已收藏!
標(biāo)簽:
保存成功

(空格分隔,最多3個(gè),單個(gè)標(biāo)簽最多10個(gè)字符)

常用:

提示

X
您的留言已提交成功!我們將在第一時(shí)間回復(fù)您~
撥打電話 產(chǎn)品分類
在線留言
青青热久免费精品视频在app| JAPAN白嫩丰满人妻VIDEOS| 欧美一区二区精品在线观看| 日韩国产欧美在线观看| 久久久久久亚洲av午夜| 亚洲日韩精品A∨片无码| 大香蕉成人在线观看| 最新日本久久中文字幕| 中文字日产幕码三区的做法大全| 欧美激情一区二区三级高清视频| 韩国理论片年轻的母亲全部| 久久久久久久三级| 国产精品美女久久久久久| 朋友的丰满人妻| 国产精品一区二区久久精品| 欧美亚洲 中文字幕| 久久综合九色综合欧美狠狠| 娇妻被交换粗又大又硬86页| 日产精品久久久久久久| 一区二区乱子伦在线播放| 中文字幕亚洲一区二区va在线| 青苹果乐园在线观看| 我要看18毛片| 久久国产亚洲AV无码麻豆| 欧洲不卡二卡三卡四卡免费| 视频黄色一区二区三区| 日韩有码中文字幕在线视频| 破了亲妺妺的处免费视频国产| 性色A∨人人爽网站HD| 调教小SAO货撅起打屁股作文| 亚洲精品国产AⅤ综合第一己| 亚洲精品中文字幕乱码三区| 黑人巨大精品欧美一区二区免费| 欧美日韩免费专区在线| 暗欲电影在线看完整| 无码专区一ⅤA亚洲V天堂| 无码乱肉视频免费大全合集| 欧美日韩大片网址| 日韩精品一区二区三区视频网| 亚洲AV成人无码久久A片| 国产精品无码久久久久久|