Derivation of endothelial cells from mammalian pluirpotent stem cells
US-2018171291-A1 · Jun 21, 2018 · US
Method for inducing vascular endothelial cells
US10669529B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10669529-B2 |
| Application number | US-201615745425-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jul 14, 2016 |
| Priority date | Jul 17, 2015 |
| Publication date | Jun 2, 2020 |
| Grant date | Jun 2, 2020 |
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Abstract
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Provided is a method for producing vascular endothelial cells from pluripotent stem cells, the method comprising the following steps (i) to (iii): (i) a step of culturing pluripotent stem cells in a culture medium comprising a BMP, on a culture vessel coated with a first matrix, to produce mesodermal progenitor cells; (ii) a step of dissociating the resulting cells into single cells; and (iii) a step of culturing the resulting cells in a culture medium comprising VEGF, on a culture vessel coated with a second matrix selected from the group consisting of laminin-411 or a fragment thereof, laminin-511 or a fragment thereof, Matrigel, type IV collagen and fibronectin.
First claim
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What is claimed is: 1. A method for producing vascular endothelial cells from pluripotent stem cells, the method comprising: (i) culturing pluripotent stem cells in a culture medium comprising a BMP, on a culture vessel coated with a first matrix, to produce mesodermal progenitor cells; (ii) dissociating the mesodermal progenitor cells obtained in the culturing of the pluripotent stem cells into single cells; and (iii) culturing the cells obtained by the dissociating of the mesodermal progenitor cells in a culture medium comprising VEGF, on a culture vessel coated with a second matrix selected from the group consisting of laminin-411 or a fragment thereof having an avidity for integrin, and, laminin-511 or a fragment thereof having an avidity for integrin. 2. The method according to claim 1 , wherein the second matrix used in the culturing of the cells obtained by the dissociating of the mesodermal progenitor cells is a fragment of laminin-411 having an avidity for integrin. 3. The method according to claim 1 , wherein the fragment of laminin-411 is laminin-411 E8. 4. The method according to claim 1 , wherein the first matrix used in the culturing of the pluripotent stem cells is Matrigel, or laminin-511 or a fragment thereof having an avidity for integrin. 5. The method according to claim 4 , wherein the fragment of laminin-511 used in culturing of the pluripotent stem cells is laminin-511 E8. 6. The method according to claim 1 , wherein the BMP is BMP4. 7. The method according to claim 1 , wherein the culture medium used in the culturing of the pluripotent stem cells further comprises a GSK3β inhibitor and VEGF. 8. The method according to claim 7 , wherein the GSK3β inhibitor is CHIR99021. 9. The method according to claim 1 , wherein the culturing of the pluripotent stem cells is carried out for two days or three days. 10. The method according to claim 1 , wherein the culturing of the cells obtained by the dissociating of the mesodermal progenitor cells is carried out for four days.
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Classifications
Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor · CPC title
Vascular endothelial growth factor [VEGF] · CPC title
Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells · CPC title
Fibronectin; Laminin · CPC title
- C12N5/069Primary
Vascular Endothelial cells · CPC title
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- What does patent US10669529B2 cover?
- Provided is a method for producing vascular endothelial cells from pluripotent stem cells, the method comprising the following steps (i) to (iii): (i) a step of culturing pluripotent stem cells in a culture medium comprising a BMP, on a culture vessel coated with a first matrix, to produce mesodermal progenitor cells; (ii) a step of dissociating the resulting cells into single cells; and (iii) …
- Who is the assignee on this patent?
- Univ Kyoto, Univ Osaka
- What technology area does this patent fall under?
- Primary CPC classification C12N5/069. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jun 02 2020 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 4 related publications on this page (citations in our corpus or others sharing the same primary CPC).