Neuro-progenitor produced from an SCNT-derived ES cell

摘要

An embryonic stem cell line derived from a nucleus-transferred oocyte prepared by transferring a nucleus of a human somatic cell into an enucleated human oocyte may differentiate into various desired cell types.

主权项

1. A method for preparing a neuro progenitor, comprising the steps of:
(1) culturing a human somatic cell to prepare a nuclear donor cell; (2) enucleating a human oocyte to prepare a recipient oocyte; (3) preparing a nucleus-transferred oocyte by transferring a nucleus of the nuclear donor cell into the recipient oocyte and fusing the nucleus of the nuclear donor cell and the recipient oocyte; (4) subjecting the nucleus-transferred oocyte to reprogramming, activation and in vitro culturing to form a blastocyst; (5) isolating an inner cell mass from the blastocyst and culturing the inner cell mass in an undifferentiated state to establish the embryonic stem cell line deposited under the accession number of KCLRF-BP-00092; (6) culturing the embryonic stem cell line to form an embryoid body; (7) culturing the embryoid body in the presence of an agent suitable for differentiating a cell of the embryoid body into the neuro progenitor; and (8) selecting a cell expressing a marker of the neuro progenitor and culturing the selected cell to obtain the neuro progenitor, wherein step (4) is performed by reprogramming the nucleus-transferred oocyte for a time period of up to 3 hours, treating the nucleus-transferred oocyte with a calcium ionophore at a concentration ranging from 6 μM to 15 μM and subsequently with 6-dimethylaminopurine at a concentration ranging from 1.5 mM to 2.5 mM, and sequentially culturing the nucleus-transferred oocyte in vitro in the G1.2 medium and the SNUnt-2 medium.