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美国Sox1-GFP报告基因小鼠胚胎干细胞的介绍
美国Sox1-GFP报告基因小鼠胚胎干细胞是美国PrimCells干细胞：PrimCells提供多种报告基因敲进的小鼠胚胎干细胞，该类细胞皆在原始基因座上敲入荧光报告基因，从而使荧光蛋白的表达能真实反映原基因座附近的表达调控状况。该类细胞作为研究胚胎干细胞自更新和定向分化的有力工具，受到多家著名研究所研究人员的青睐。
| Species: | Mus Musculus | Catalog Number: | PCEMM01 |
| Cell Number: | ~2x106 | Storage/Shipping: | Dry Ice |
Introduction:
Sox1 is a transcriptional factor from SOX protein family. It is one of the earliest and most specific markers for neural progenitors and expressed throughout the central neural system during early embryo development1, 2. PrimCells is proud to isolate and provide to the research community the culture of Sox1-EGFP ESCs from transgenic knock-in mice. These cells were cultured in standard ESCs medium on gelatin coated plates and be used as a powerful tool to examine the process by which mESCs acquire neural identity.
ES cell culture and monolayer neural differentiation procudures
Sox1-GFP mESCs should be maintained in the stem cell culture medium to keep the pluripotent state (Figure 1).
For monoculture neural differentiation, dissociate and plate the undifferentiated Sox1-GFP mESCs on to 0.1% gelatin-coated tissue culture plastic at a density of 1-3x104/cm2 in serum-free N2B27 medium. Renew N2B27 medium every 2 days. GFP positive cells can be observed on Day 5 (Figure 2).
Figure 1 | Sox1-GFP mESCs express typical ES cell markers
Figure 2 | Monoculture neuronal differentiation of Sox1-GFP mESCs on Day 5
Growth Medium
Embryonic stem cell medium
DMEM: 415ml
FBS: 75ml (HYCLONE LABORATORIES INC, Cat# SH30070.03EH)
L-Glutamine: 5ml
MEM NEAA: 5ml
1-Thioglycerol: 4.3ul
LIF: 50ul LIF: 50ul (10ng/ml final concentration) (PrimCells, Cat# PFHSA01)
Total Volume: 500ml
N2B27 medium for neural differentiation
N2B27 is a 1:1 mixture of DMEM/F12 (Gibco, Paisley, UK) supplemented with modified N2 (25 µg/ml insulin, 100 µg/ml apo-transferrin, 6 ng/ml progesterone, 16 µg/ml putrescine, 30 nM sodium selenite and 50 µg/ml bovine serum albumin fraction V (Gibco)) and Neurobasal medium supplemented with B27 (both from Gibco).
Alternatively, Ndiff227 (StemCells INC, Cat# SCS-SF-NB-02) is ready-to-use medium for the neural differentiation of mESCs in adherent monoculture conditions as described in Ying QL et al (2003)2.
References
1. Aubert, J. et al. Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice. Proc Natl Acad Sci U S A 100 Suppl 1, 11836-11841 (2003).
2. Ying, Q.L., Stavridis, M., Griffiths, D., Li, M. & Smith, A. Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. Nat Biotechnol 21, 183-186 (2003).
美国PrimCells品牌介绍
PrimCells是位于美国加州圣迭戈的一家专业致力于细胞产品研发和解决方案提供的业内技术。
公司汇集了资深的生物医药技术专家，独立开发了多项原代细胞和干细胞相关的专有技术。 公司以“细胞研究的优选择"为宗旨，自主研发超过70种创新产品，多个特种细胞市场的空白。例如其诱导性表达细胞库和高效3D皮肤培养系统，皆为该类产品目前市场上少数的规?；┯ι?。此外，公司以高质量的数据与快速稳定的项目完成时间，长期为多家制药公司提供解决方案，获得用户广泛赞誉。
PrimCells产品线：
报告基因干细胞 2. 原代细胞（含报告基因原代细胞） 3. 诱导性稳定细胞株
4. 人源重组蛋白 5. 创新型培养试剂
PrimCells 解决方案
1.诱导性稳定细胞株开发 2. 原代细胞开发 3. 药物体外毒理和效能测试 4. 3D-皮肤培养
PrimCells合作伙伴及客户:
A．生物制药公司：辉瑞(Pfizer), 诺华(Novartis), 礼来(Eli2 Lilly),. Amgen, EMD Millipore, Regeneron等公司的毒理和医药研发部门
B．科研机构：冷泉港实验室，哈佛医学院, 麻省理工（MIT）,， UCSF, NYU, Salk, UIUC, USC等
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