	US 12,186,345 B2
	Cell population including adhesive stem cells, production method therefor and pharmaceutical composition
Nobuyoshi Umeda, Kobe (JP); Keita Ino, Kobe (JP); and Chiho Kobayashi, Kobe (JP)
Assigned to KANEKA CORPORATION, Osaka (JP)
Appl. No. 16/957,896
Filed by KANEKA CORPORATION, Osaka (JP)
PCT Filed Dec. 28, 2018, PCT No. PCT/JP2018/048540 § 371(c)(1), (2) Date Jun. 25, 2020, PCT Pub. No. WO2019/132026, PCT Pub. Date Jul. 4, 2019.
Claims priority of application No. 2017-253879 (JP), filed on Dec. 28, 2017.
Prior Publication US 2020/0368291 A1, Nov. 26, 2020
Int. Cl. A61K 35/545 (2015.01); C12N 5/0775 (2010.01)

CPC A61K 35/545 (2013.01) [C12N 5/0665 (2013.01)]

9 Claims

1. A method for producing a cell population comprising adherent stem cells, comprising:

obtaining fetal membrane and placenta from a female patient;

manually separating amnion from the stump of the fetal membrane;

enzymatically treating the amnion comprising epithelial cell layer and adherent stem cell layer to collect amniotic adherent stem cells;

culturing the enzymatically treated and collected amniotic adherent stem cells derived from a fetal appendage in a medium comprising human platelet lysate (hPL) and obtaining a cultured cell population;

measuring a proportion of potassium channel, voltage gated subfamily A regulatory beta subunit 1-positive (KCNAB1-positive) of the cultured cell population;

identifying a cell population comprising adherent stem cells maintaining a normal karyotype by using the proportion of KCNAB1-positive adherent stem cells being 85% or more in as an index to obtain an identified cell population; and

obtaining the identified cell population identified by using said index which comprise adherent stem cells maintaining a normal karyotype by using the proportion of KCNAB1-positive adherent stem cells being 85% or more.