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  • Efficient derivation of extraembryonic endoderm stem cell lines from mouse postimplantation embryos.

Efficient derivation of extraembryonic endoderm stem cell lines from mouse postimplantation embryos.

Scientific reports (2016-12-20)
Jiangwei Lin, Mona Khan, Bolek Zapiec, Peter Mombaerts
ABSTRACT

Various types of stem cell lines have been derived from preimplantation or postimplantation mouse embryos: embryonic stem cell lines, epiblast stem cell lines, and trophoblast stem cell lines. It is not known if extraembryonic endoderm stem (XEN) cell lines can be derived from postimplantation mouse embryos. Here, we report the derivation of 77 XEN cell lines from 85 postimplantation embryos at embryonic day E5.5 or E6.5, in parallel to the derivation of 41 XEN lines from 69 preimplantation embryos at the blastocyst stage. We attain a success rate of 100% of XEN cell line derivation with our E5.5 whole-embryo and E6.5 disaggregated-embryo methods. Immunofluorescence and NanoString gene expression analyses indicate that the XEN cell lines that we derived from postimplantation embryos (post-XEN) are very similar to the XEN cell lines that we derived from preimplantation embryos (pre-XEN) using a conventional method. After injection into blastocysts, post-XEN cells contribute to extraembryonic endoderm in chimeras at E6.5 and E7.5.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Heparin sodium salt from porcine intestinal mucosa, Grade I-A, ≥180 USP units/mg, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
M2 medium, With HEPES, without penicillin and streptomycin, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
Tyrode′s Solution, Acidic, liquid, sterile-filtered, suitable for mouse embryo cell culture