美國(guó)Sox1-GFP報(bào)告基因小鼠胚胎干細(xì)胞的介紹

美國(guó)Sox1-GFP報(bào)告基因小鼠胚胎干細(xì)胞是美國(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，受到多家著名研究所研究人員的青睞。

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 development^{1, 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)。

公司匯集了資深的生物醫(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)胞研究的優(yōu)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)?；?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:

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