美國(guó)Sox1-GFP報(bào)告基因小鼠胚胎干細(xì)胞
美國(guó)PrimCells干細(xì)胞：PrimCells提供多種報(bào)告基因敲進(jìn)的小鼠胚胎干細(xì)胞，該類(lèi)細(xì)胞皆在原始基因座上敲入熒光報(bào)告基因，從而使熒光蛋白的表達(dá)能真實(shí)反映原基因座附近的表達(dá)調(diào)控狀況。該類(lèi)細(xì)胞作為研究胚胎干細(xì)胞自更新和定向分化的有力工具，受到多家*研究所研究人員的青睞。
| 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).
美國(guó)PrimCells品牌介紹
PrimCells是位于美國(guó)加州圣迭戈的一家專(zhuān)業(yè)致力于細(xì)胞產(chǎn)品研發(fā)和解決方案提供的業(yè)內(nèi)技術(shù)。
公司匯集了*的生物醫(yī)藥技術(shù)專(zhuān)家，獨(dú)立開(kāi)發(fā)了多項(xiàng)原代細(xì)胞和干細(xì)胞相關(guān)的專(zhuān)有技術(shù)。 公司以“細(xì)胞研究的*選擇”為宗旨，自主研發(fā)超過(guò)70種創(chuàng)新產(chǎn)品，*多個(gè)特種細(xì)胞市場(chǎng)的空白。例如其誘導(dǎo)性表達(dá)細(xì)胞庫(kù)和高效3D皮膚培養(yǎng)系統(tǒng)，皆為該類(lèi)產(chǎn)品目前市場(chǎng)上少數(shù)的規(guī)模化供應(yīng)商。此外，公司以高質(zhì)量的數(shù)據(jù)與快速穩(wěn)定的項(xiàng)目完成時(shí)間，長(zhǎng)期為多家制藥公司提供解決方案，獲得用戶(hù)廣泛贊譽(yù)。
PrimCells產(chǎn)品線：
- 報(bào)告基因干細(xì)胞 2. 原代細(xì)胞（含報(bào)告基因原代細(xì)胞） 3. 誘導(dǎo)性穩(wěn)定細(xì)胞株
4. 人源重組蛋白 5. 創(chuàng)新型培養(yǎng)試劑
PrimCells 解決方案
1.誘導(dǎo)性穩(wěn)定細(xì)胞株開(kāi)發(fā) 2. 原代細(xì)胞開(kāi)發(fā) 3. 藥物體外毒理和效能測(cè)試 4. 3D-皮膚培養(yǎng)
PrimCells合作伙伴及客戶(hù):
A．生物制藥公司：輝瑞(Pfizer), 諾華(Novartis), 禮來(lái)(Eli2 Lilly),. Amgen, EMD Millipore, Regeneron等公司的毒理和醫(yī)藥研發(fā)部門(mén)
B．科研機(jī)構(gòu)：冷泉港實(shí)驗(yàn)室，哈佛醫(yī)學(xué)院, 麻省理工（MIT）,， UCSF, NYU, Salk, UIUC, USC等
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