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。

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PrimCells品牌介紹

PrimCells是位于美國加州圣迭戈的一家專業致力于細胞產品研發和解決方案提供的業內技術。

公司匯集了資深的生物醫藥技術專家，獨立開發了多項原代細胞和干細胞相關的專有技術。 公司以“細胞研究的優選擇"為宗旨，自主研發超過70種創新產品，多個特種細胞市場的空白。例如其誘導性表達細胞庫和高效3D皮膚培養系統，皆為該類產品目前市場上少數的規模化供應商。此外，公司以高質量的數據與快速穩定的項目完成時間，長期為多家制藥公司提供解決方案，獲得用戶廣泛贊譽。

PrimCells產品線：

• 報告基因干細胞        2. 原代細胞（含報告基因原代細胞）               3. 誘導性穩定細胞株

4. 人源重組蛋白               5.   創新型培養試劑

PrimCells 解決方案

1.誘導性穩定細胞株開發 2. 原代細胞開發 3. 藥物體外毒理和效能測試 4. 3D-皮膚培養

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等公司的毒理和醫藥研發部門

B．科研機構：冷泉港實驗室，哈佛醫學院, 麻省理工（MIT）,， UCSF, NYU, Salk, UIUC, USC等