性做久久久久久坡多野结衣-性做久久久久久久久浪潮-性欲影院-性影院-国产精品线路一线路二-国产精品兄妹在线观看麻豆

武漢原生原代生物醫藥科技有限公司

PriCells: Primary brain cell isolation and culture.

時間:2022-1-18 閱讀:283
分享:

PriCells: Primary brain cell isolation and culture.                                                      


1. Cerebella were removed from 7-day-old mice and passed through Nitex nylon netting (80 μm pore size) into primary cell system containing 20% (v/v) fetal calf serum (FCS).

2. The cell suspension was plated at a density of four cerebella per 12-well culture plate.

3. Primary cell culture system was changed two days after plating and twice a week thereafter, gradually decreasing the FCS concentration to 10%.

4. At 14 days in culture, dibutyryl-cAMP was added to the medium for 1 week to promote the morphological differentiation of astrocytes.

5. Experiments were performed on 3-week-old cultures.

6. Cortical astrocytes were prepared from 1-day-old mice following the same procedure used for cerebellar astrocytes plated at a density of two cortices per 12-well culture plate.

7. Cerebellar neurons were isolated and cultured from the cerebellum of 7-day-old mice, after mild trypsinization of the tissue followed by trituration in a DNase solution containing a trypsin inhibitor derived from soybeans.

8. Cells were suspended in a slightly modified primary cell culture containing 50 μM kainic acid and 10% (v/v) FCS and plated at density 2 × 106 cells per well in a 12-well culture plate.

9. The wells had been coated with poly-L-lysine.

10. Cytosine arabinoside (20 μM) was added after 48 h to prevent astrocyte proliferation.

11. Cells were used for experiments after 1 week in culture.

12. For the co-cultures, the neurons and astrocytes were prepared separately and seeded as described earlier.

13. The astrocytes were cultured on inserts for 21 days, and the neurons were cultured in culture plates for 7 days prior to incubation in co-culture.

14. The astrocytes were grown on permeable membranes in tissue culture plate inserts, and the neurons were cultured in standard tissue culture plates.

15. The inserts were transferred to the neuronal plates 24 h prior to incubation.

16. At time of insert transfer, half of the neuronal medium was extracted and replaced with astrocyte cell culture system.

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: 佛坪县| 双江| 左云县| 大理市| 康定县| 濮阳县| 卫辉市| 蓬莱市| 黑水县| 大安市| 肥西县| 惠水县| 阳西县| 拉萨市| 宝清县| 盐山县| 三原县| 涟源市| 珠海市| 兰溪市| 唐海县| 石首市| 江北区| 江西省| 宁蒗| 新巴尔虎左旗| 平安县| 蓬安县| 湘潭县| 龙井市| 乐东| 普定县| 城市| 乌鲁木齐县| 文水县| 罗江县| 南江县| 兴山县| 北安市| 什邡市| 金堂县|