In vitro maturation culture medium of immature oocytes and use thereof
AssigneeChina Agricultural University
InventorsGuoliang Xia, Chao Wang, Huarong Wang, Han Cai
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Abstract
Provided is a method for culturing immature oocytes. The method can promote in vitro maturation of the immature oocytes, and specifically comprises using follicular cells and a culture medium for culturing same. The culture medium for culturing the follicular cells contains CNP or variants thereof or analogues thereof and an HDAC (histone deacetylase) inhibitor. Also provided are the in vitro maturation culture medium containing CNP or variants thereof or analogues thereof and the HDAC inhibitor, and related compositions thereof, and the use of the above medium, culture medium and compositions in the promotion of in vitro maturation of the immature oocytes.
Claims12
A culture medium, comprising follicular granule cells and a culture solution for culturing the follicular granule cells, wherein the culture solution for culturing the follicular granule cells comprises 30˜120 ng/mL of C-type natriuretic peptide (CNP) and 1-10 μM of a histone deacetylase (HDAC) inhibitor, wherein the HDAC inhibitor is a HDAC3 inhibitor selected from the group consisting of HDACi 4b, Entinostat (MS-275), BG45, RG2833 (RGFP109) and RGFP966, or a mixture thereof.
A culture solution for in vitro maturation of an immature oocyte, comprising 30-120 ng/mL of CNP and 1-10 μM of a HDAC inhibitor, and is conditioned through contacting the culture solution with follicular granule cells, wherein the HDAC inhibitor is a HDAC3 inhibitor selected from the group consisting of HDACi 4b, Entinostat (MS-275), BG45, RG2833 (RGFP109) and RGFP966, or a mixture thereof.
A method for promoting the in vitro maturation of immature oocytes, comprising, collecting granule cells from a follicle of a subject, culturing the granule cells in a cell monolayer adherent culture in a culture solution comprising 30-120 ng/mL of CNP and 1-10 μM of a HDAC3 inhibitor to obtain feeder layer cells, collecting oocytes in the germinal vesicle stage from the subject, and culturing the oocytes in the culture solution containing the feeder layer cells for 20˜25h, wherein the HDAC inhibitor is a HDAC3 inhibitor selected from the group consisting of HDACi 4b, Entinostat (MS-275), BG45, RG2833 (RGFP109) and RGFP966, or a mixture thereof.
Dependencies are inferred from the claim text.