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美國Sox1-GFP報告基因小鼠胚胎干細胞的介紹
美國Sox1-GFP報告基因小鼠胚胎干細胞是美國PrimCells干細胞：PrimCells提供多種報告基因敲進的小鼠胚胎干細胞，該類細胞皆在原始基因座上敲入熒光報告基因，從而使熒光蛋白的表達能真實反映原基因座附近的表達調控狀況。該類細胞作為研究胚胎干細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PrimCells品牌介紹(shao)
PrimCells是位于美國加州圣迭戈的一家專業(ye)致(zhi)力于細胞產品研發和解決方案提供的業(ye)內技(ji)術(shu)。
公司匯(hui)集了資(zi)深的(de)(de)生物醫(yi)藥(yao)技(ji)術專家，獨立開發了多項原代(dai)細胞(bao)和干細胞(bao)相關(guan)的(de)(de)專有技(ji)術。 公司以“細胞(bao)研(yan)究的(de)(de)優選(xuan)擇"為(wei)宗(zong)旨，自主研(yan)發超過(guo)70種創新產品，多個特種細胞(bao)市場的(de)(de)空白。例如(ru)其誘導(dao)性表達細胞(bao)庫(ku)和高(gao)效(xiao)3D皮膚(fu)培養系統，皆為(wei)該類產品目前市場上(shang)少數(shu)的(de)(de)規模化供應商。此外(wai)，公司以高(gao)質量的(de)(de)數(shu)據與快速穩定(ding)的(de)(de)項目完成(cheng)時間，長期為(wei)多家制藥(yao)公司提(ti)供解決方案，獲得用戶(hu)廣泛贊譽。
PrimCells產品線(xian)：
報告(gao)基因干細胞(bao) 2. 原(yuan)代細胞(bao)（含報告(gao)基因原(yuan)代細胞(bao)） ; 3. 誘(you)導性穩定細胞(bao)株
4. 人(ren)源(yuan)重組蛋白 5. 創(chuang)新(xin)型(xing)培養試劑
PrimCells 解決(jue)方案(an)
1.誘導(dao)性穩定細(xi)胞株開(kai)發 2. 原代細(xi)胞開(kai)發 3. 藥物(wu)體外毒理和效(xiao)能測試 4. 3D-皮膚(fu)培養
PrimCells合作伙(huo)伴及客戶:
A．生物制藥(yao)公司：輝瑞(Pfizer), 諾華(Novartis), 禮來(Eli2 Lilly),. Amgen, EMD Millipore, Regeneron等公司的(de)毒理(li)和醫(yi)藥(yao)研發部門(men)
B．科研(yan)機構：冷泉港實驗室，哈佛醫學院, 麻省理工（MIT）,， UCSF, NYU, Salk, UIUC, USC等
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