人雪旺细胞描述：
雪旺细胞是神经嵴的衍生物，形成了周围神经髓磷脂轴突的髓鞘。它环绕在末梢神经轴突的轴上，沿着轴突节形成一层髓磷脂髓鞘。雪旺细胞对末梢神经的发育、功能和再生起着重要作用。当轴突快死亡时，雪旺细胞环绕其周围帮助消化轴突。连续的雪旺细胞形成一个空的管道，在其尾端形成新的轴突。末梢神经中的雪旺细胞的数量是受到严格调控的。体外增殖受到诸如PDGF, FGF，神经元和其它多肽类生长因子的刺激。雪旺细胞为研究一系列发育问题提供了相对简单、明确、易懂的哺乳动物模型。在研究神经病和神经再生，以及中枢神经系统修复方面，雪旺细胞也具有特殊的临床重要性。
人雪旺细胞 (HSC)提取于人脊神经组织，原代冻存。每管含有细胞数>5×105 cells/ml，此细胞通过对S-100, GFAP和CD90免疫荧光染色验证，经测试不含有HIV-1、HBV、HCV、支原体、细菌、酵母和真菌。
配套培养基: 雪旺细胞培养基(SCM, Cat. No. 1701)
产品使用说明：本产品仅用于科研研究使用1.) Lin, G., Zhang, H., Sun, F., Lu, Z., Reed-Maldonado, A., Lee, Y.C., Wang, G., Banie, L. & Lue, T.F. (2016) Brain-derived neurotrophic factor promotes nerve regeneration by activating the JAK/STAT pathway in Schwann cells Translational Andrology and Urology. 5
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Question:1
What is the best way you suggest to cryopreserve primary human Schwann cells?
Answers:
Posted by Karishma Shah on Friday, March 4, 2016
Dear customer,
We do not recommend that customers re-freeze our cells since primary cells are fragile and they may be damaged during re-freeze and re-thaw process.
Question:2
In the product description, these cells were analyzed by immunofluorescence using antibodies against S100, GFAP, and CD90. Would you be able to provide manufacturer/catalog# for those antibodies? Your protocols for cell preparation and immunofluorescence using those antibodies would be kindly appreciated as well? Thank you
Answers:
Posted by Manoj Sharma on Thursday, January 14, 2016
Dear Amish,
Here are the antibodies we use for staining #1700 Human Schwann Cells:
1. S-100 beta:
Sigma
Cat. No. S2532
1:500 dilution
2. GFAP:
Sigma
Cat. No. G3893
1:1000 dilution
And following is the protocol that we use for staining the cells.
Immunofluorescence Staining Protocol
a. Fixing the slide
i. Wash each well with 500 μl PBS making sure to add along the side of the well wall.
ii. Add 500 μl 4% PFA (paraformaldehyde) to each well and place at room temperature for 5 minutes.
b. Blocking Solution
i. Wash each well with 500 μl PBS.
ii. The blocking solution consists of 5% NGS (Normal Goat Serum) and 0.1% Triton-X-100 in PBS.
iii. The blocking solution can be made in bulk in a 15mL conical tube and then placed in the wells.
iv. For every 1 ml of PBS add 50 μl of NGS and 10 μl of 10% Triton-X 100. Add 250 μl per well and incubate at room temperature for 1 hour.
c. Primary Antibody
i. Prepare the solution for the primary antibody. This consists of 1% NGS and 1% Triton-X 100 in PBS. For every 1 ml of PBS add 10 μl of NGS and 10 μl of Triton-X 100. This can be made in bulk in a conical tube.
ii. Dilute each primary antibody needed with the necessary dilution factor (depending on the antibody used) and add 250 μl to the appropriate wells. Place the slide in 4°C overnight. Dilutions can be made in 1.5mL microcentrifuge tubes.
iii. If the slide needs be stained on the same day, it can be placed in the 37°C incubator for 2 hours instead of 4°C overnight.
iv. Make sure to note on the Immunocytochemistry form the specific antibody placed in the well.
d. Secondary Antibody
i. Wash the slide 3 times with 500 μl PBS leaving the PBS in the wells 5-7 minutes per rinse.
ii. Dilute each secondary antibody (usually 1:1000 to 1:2000, depending on the intensity) needed with PBS and add 250 μl to the appropriate wells. Incubate at 37°C in the incubator for 45 minutes.
iii. Wash the slide 5 times with 500 μl PBS leaving the PBS in the wells 5-7 minutes per rinse.
iv. The secondary antibody is specific to the primary antibody such as IgG Rabbit or IgG Mouse. The animal it derives from must be the same as the primary antibody.
e. Preparing the slide for microscopic examination
i. Drop about 5-8 μl of mounting medium (DAPI) to a slide, remove cover slips from the well using tweezers and place them over the slide on the mounting medium gently.
ii. Make sure the DAPI covers the entire area of the cover slips.
f. Microscopic Examination
Examine the slides after 20-30 minutes under fluorescence microscope.
Thank you for your interest in ScienCell Products
Manoj
Question:3
Has anyone used these cells to look at myelin production? I work with a virus that may disrupt myelination and I was interested in looking at this in primary tissue culture cells. Thanks! Lee
Answers:
Posted by Manoj Sharma on Thursday, January 7, 2016
Hi Lee,
We do not test for myelin production.
thank you for your interest.
SCIENCELL TEAM
Question:4
can you send an instruction on how to culture them for a few passages before we can use them for treatments? Thanks, tao
Answers:
Posted by Karishma Shah on Tuesday, October 6, 2015
Dear Tao,
Thanks for asking! Culture information is located on the product sheet on the product page under "Technical Resources."
For your convenience, here is the product sheet for #1700: http://sciencellonline.com/downloads/dl/file/id/249/1700.pdf
Thanks!
上海中乔新舟生物科技有限公司
