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Growth of embryonic stem cells

A technology of embryonic stem cells and embryos, applied in the field of cell biology, can solve the problems of time-consuming, complex and labor-intensive

Inactive Publication Date: 2008-02-20
MILLIPORE CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Therefore, the conditions for culturing and maintaining ESCs from any source in vitro are generally complicated, time-consuming, labor-intensive, difficult and expensive.

Method used

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  • Growth of embryonic stem cells
  • Growth of embryonic stem cells
  • Growth of embryonic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: fibronectin coating method

[0045] Material:

[0046] Fibronectin 0.1% solution (Sigma, St Louis, MO)

[0047] Untreated primary mouse embryonic fibroblasts (MEFs) of the CF-1 line (Chemicon, Temecula, CA)

[0048] MEF medium: DMEM (Invitorgen, Carlsbad, CA)

[0049] 10% fetal bovine serum (Hyclone Logan, UT)

[0050] 1% Glutamax-1 (Invitorgen, Carlsbad, CA)

[0051] 1% PenStrep (Invitorgen, Carlsbad, CA)

[0052] 1% non-essential amino acids (Sigma, St Louis, MO)

[0053] DPBS (Hyclone Logan, UT)

[0054] Fibronectin (0.1% solution) (Sigma, St Louis, MO) was diluted to 25 μg / ml in sterile DPBS. Sufficient amount of fibronectin (25 μg / ml) in DPBS was added to coat the one-well tray (approximately 5-10 mL / dish). Incubate the tray at room temperature for at least 45 min. Excess fibronectin was removed prior to inoculation of MEFs. Thaw MEFs stored in a -80°C freezer. Gently shake the MEF bottle in a 37 °C water bath. Transfer MEFs to a 15 mL tub...

Embodiment 2

[0055] Example 2: Indirect co-cultivation of embryonic stem cells and feeder cells

[0056] Materials used:

[0057] Millipore Cell Culture Filter Plates and Single Well Feeder Trays, 24 Wells, with 0.4 μm PCF Membrane or 1.0 μm PET Membrane

[0058] 96-well 0.4 μm PCF membrane (Millipore Corp., Billerica, MA)

[0059]ESC Medium: Knock out DMEM (Invitorgen, Carlsbad, CA)

[0060] 20% ES certified serum (Invitorgen, Carlsbad, CA)

[0061] 1% Glutamax-1 (Invitorgen, Carlsbad, CA)

[0062] 1% PenStrep (Invitorgen, Carlsbad, CA)

[0063] 1% non-essential amino acids (Sigma, St Louis, MO)

[0064] 0.1% ESGRO(R) (LIF) (Chemicon, Temecula, CA)

[0065] 0.1% 2-mercaptoethanol (Invitorgen, Carlsbad, CA)

[0066] MEF medium:

[0067] DMEM (Invitorgen, Carlsbad, CA)

[0068] 10% fetal bovine serum (Hyclone Logan, UT)

[0069] 1% Glutamax-1 (Invitorgen, Carlsbad, CA)

[0070] 1% PenStrep (Invitorgen, Carlsbad, CA)

[0071] 1% non-essential amino acids (Sigma, St Louis, MO)

...

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Abstract

The invention provides systems, kits and methods relating to the growth of stem cells.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application No. 60 / 816,514, filed June 26, 2006, which is hereby incorporated by reference in its entirety. technical field [0003] The present invention relates generally to the field of cell biology. In certain specific embodiments, the present invention provides systems, kits and methods related to the growth of stem cells. Background technique [0004] Embryonic stem cells (ESCs) can be derived from pre-embryonic, embryonic or fetal tissue at any time after fertilization. The isolation and cultivation of various types of stem cells has been described previously, see for example Robertson, 1997, Methods of Cell Biology 75:173; Thompson et al.1995, Proc.Natl.Acad.Sci.USA 92:7844; Thompson et al .1998,Science 282:114;美国专利No.5,166,065;5,332,672;5,405,772;5,453,357;5,639,618;5,672,499;5,843,780;5,914,268;5,922,597;5,968,829;6,040,180;6,090,622;6,833,269;6,506,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/0735
CPCC12N2533/30C12N2502/02C12N5/0606C12N2502/13A61P17/02A61P31/00A61P37/02A61P39/00A61P43/00
Inventor S·D·谢里登S·吉尔
Owner MILLIPORE CORP
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