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Functional Genomics & RNA
Trophoblast Protocol
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The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in an exclusive partnership with Sigma-Aldrich. Each month, 2 new protocols will be featured, each with a list of products available from Sigma-Aldrich.
View the entire protocol on BioSupplyNet.com.
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View the entire protocol "Culturing Trophoblast Stem (TS) Cell Lines" at BioSupplynet.com
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Janet Rossant
This protocol was adapted from "Isolation and Culture of Blastocyst-Derived Stem Cell Lines," Chapter 8 (Protocol 8, provided by the laboratory of Janet Rossant), in Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.
ABSTRACT
Recently, culture conditions have been established for a second blastocyst-derived cell line, trophoblast stem (TS) cells, in addition to embryonic stem (ES) cells. This protocol describes a method for culturing TS cell lines. These cells can then be used to study trophoblast differentiation and placental function. |
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| Products Available for this Protocol |
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Dimethylsulfoxide |
D2650 |
Dimethyl sulfoxide, liquid (temperature dependent), ≥99.7%, Hybri-Max™, sterile-filtered, hybridoma tested |
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| Heparin |
H4784 |
Heparin sodium salt, ~170 USP units/mg |
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| Gelatin (Swine Skin) |
G2500 |
Gelatin from porcine skin, Type A |
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| Trypsin-EDTA |
T3924 |
Trypsin-EDTA solution, 1 ×, 0.5 g porcine trypsin and 0.2 g EDTA • 4Na per liter of Hanks' Balanced Salt Solution with phenol red., sterile-filtered, cell culture tested |
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| BSA fraction V |
A4503 |
Albumin from bovine serum, Cohn V fraction, ≥96% (agarose gel electrophoresis), lyophilized powder |
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| PBS |
P5368 |
Phosphate buffered saline, pH 7.4, Biotechnology Performance Certified |
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| Mitomycin C stock |
M4287 |
Mitomycin C from Streptomyces caespitosus, powder, cell culture tested |
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| Dulbecco’s modified Eagle’s medium (DMEM) |
D6046 |
Dulbecco’s Modified Eagle’s Medium - low glucose, With 1000 mg/L glucose, L-glutamine, and sodium bicarbonate, liquid, sterile-filtered, cell culture tested |
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| Fetal bovine serum (FBS) |
F2442 |
Fetal Bovine Serum, USA origin, sterile-filtered, cell culture tested, hybridoma tested |
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| Sodium pyruvate |
P5280 |
Sodium pyruvate, cell culture tested, insect cell culture tested, ≥99%, powder |
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| β-mercaptoethanol |
M3148 |
2-Mercaptoethanol, for molecular biology, ≥98% (GC/titration) |
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| L-glutamine |
G6392 |
L-Glutamine, γ-irradiated, cell culture tested |
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| Penicillin and Streptomycin |
P4333 |
Penicillin-Streptomycin, 100 ×, liquid, stabilized, sterile-filtered, cell culture tested |
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| RPMI 1640 |
R8758 |
RPMI-1640 Medium, with L-glutamine and sodium bicarbonate, liquid, sterile-filtered, cell culture tested |
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Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.
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