Embryonic cell cultures and methods of using the same

Patent metadata

Field	Value
Publication number	US-11913015-B2
Application number	US-201816606199-A
Country	US
Kind code	B2
Filing date	Apr 17, 2018
Priority date	Apr 17, 2017
Publication date	Feb 27, 2024
Grant date	Feb 27, 2024

Abstract

The invention provides a composition comprising an extraembryonic endodermal (XEN) call and/or an embryonic fibroblast (EF) cell. The invention also provides a method of establishing a XEN cell line or a primary embryonic fibroblast (EF) cell line in vitro, the method comprising culturing a zygote or parthenote from a mammal for a time sufficient to produce one or more blastocysts; and culturing the one or more blastocysts on feeder cells in culture medium for a time sufficient to produce one or a plurality of XEN cells and/or one or a plurality of EF cells.

First claim

The invention claimed is:

1. A composition comprising an isolated extraembryonic endodermal (XEN) cell;
wherein the isolated XEN cell is a porcine cell; and
wherein the isolated XEN cell comprises a phenotype of CDX2−, NANOG−, SOX2−, GATA4+, FOXA2+, GATA6+, SALL4+ and SOX17+.

2. The composition of claim 1 , wherein the isolated XEN cell is derived from a blastocyst outgrowth, wherein the blastocyst outgrowth was cultured for a duration of about 4 days to about 6 days.

3. The composition of claim 1 , wherein the isolated XEN cell is derived from an epiblast.

4. The composition of claim 1 , wherein the isolated XEN cell comprises a genetic modification.

5. The composition of claim 1 , wherein the composition is free or substantially free of one or a combination of: (i) embryonic epiblastic cells; (ii) cells from a fetus of an animal; and (iii) trophoblastic cells.

6. A library comprising at least two different isolated cell lines, wherein one of the at least two isolated cell lines is an extraembryonic endodermal (XEN) cell line, wherein the XEN cell line is a porcine cell line and wherein the XEN cell line comprises a phenotype of CDX2−, NANOG−, SOX2−, GATA4+, FOXA2+, GATA6+, SALL4+ and SOX17+.

7. The library of claim 6 , wherein each of the at least two different isolated cell lines has a different genetic modification.

8. The composition of claim 1 , wherein the isolated XEN cell is in culture for at least about 5 days.

9. The composition of claim 1 , wherein the isolated XEN cell is passaged greater than 40 times.

10. The composition of claim 4 wherein the genetic modification is a knock-in, knock-out, point mutation or deletion.

11. The composition of claim 1 , wherein the isolated XEN cell further comprises a phenotype of EOMES+.

Assignees

• Univ Maryland

Inventors

• Park Chi-Hun
• Telugu Bhanu Prakash V L

Classifications

• C12N15/877 Primary

  Techniques for producing new mammalian cloned embryos · CPC title

• A01K67/027

  New or modified breeds of vertebrates · CPC title

• C12N5/0605 Primary

  Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly · CPC title

• C12N2510/00

  Genetically modified cells · CPC title

• C12N2830/008

  cell type or tissue specific enhancer/promoter combination · CPC title