美國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，受到多家有名研究所研究人員的青睞。
| Species: | Mus Musculus | Catalog Number: | PCEMM01 |
| Cell Number: | ~2x106 | Storage/Shipping: | Dry Ice |
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 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PrimCells是位于美國加州圣迭戈的一家專業(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)領(lǐng)導(dǎo)企業(yè)。
公司匯集了**的生物醫(yī)藥技術(shù)專家，獨(dú)立開發(fā)了多項(xiàng)原代細(xì)胞和干細(xì)胞相關(guān)的專有技術(shù)。 公司以“細(xì)胞研究的選擇”為宗旨，自主研發(fā)超過70種**產(chǎn)品，填補(bǔ)了多個(gè)特種細(xì)胞市場的空白。例如其誘導(dǎo)性表達(dá)細(xì)胞庫和高效3D皮膚培養(yǎng)系統(tǒng)，皆為該類產(chǎn)品目前市場上少數(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，長期為多家知名制藥公司提供解決方案，獲得用戶廣泛贊譽(yù)。
PrimCells產(chǎn)品線：
PrimCells 解決方案
1.誘導(dǎo)性穩(wěn)定細(xì)胞株開發(fā) 2. 原代細(xì)胞開發(fā) 3. **體外毒理和效能測試 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(Eli2 Lilly),. Amgen, EMD Millipore, Regeneron等公司的毒理和醫(yī)藥研發(fā)部門
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
