A kind of endometrial organoid culture medium and culture method

Abstract

The invention provides an endometrial organoid culture medium and a culturing method. The culture medium comprises, according to the final concentration, EGF at 10-100ng / ml, Noggin at 20-500ng / ml, and R-spondin at 20-500ng / ml. 1, 20‑500ng / ml Wnt3a, 1‑100ng / ml FGF9, 10‑200ng / ml KIAA1199, 1‑100ng / ml sox2, 10‑100nM isoflavones, 100‑2000nM CHIR99021, 1‑40µM A83‑01, 2‑50μM ROCK inhibitor, 50‑200ug / ml primary cell antibiotic; the above components are all dissolved in DMEM / F12 medium. This medium is used for endometrial organoid culture, which is conducive to the rapid expansion of endometrial stem cells and the maturation and differentiation of organoids, and the number of passages is significantly increased, and the same tissue as the primary tissue can still be maintained after multiple passages. Structural and genomic properties.

Description

technical field [0001] The invention relates to the technical field of organoid culture, in particular to an endometrial organoid culture medium and a culture method. Background technique [0002] The endometrium undergoes a process of dynamic development. During reproduction, the functional layers of the endometrium undergo regeneration, differentiation, and shedding during the menstrual cycle under the control of the hypothalamic-pituitary-ovarian axis. The mucosa contains a single layer of glands lined by a secretory columnar epithelium separated by a stroma. The basic functions of the endometrium are the luminal and glandular epithelium, consisting of a mixture of cilia and secretory cells. The luminal epithelium is the site of embryo attachment, the epithelium covers the surface of the endometrium and invaginates from the endometrium to the stroma to form glands. Glandular secretions are rich in growth factors and lipids necessary for placental growth. [0003] Organ...

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Problems solved by technology

[0004] Although a variety of human-derived other tissues (such as liver, intestine, etc.) can be successfully cultured into organoids in vitro using different methods and under different culture conditions, the current research and reports on the culture methods of endometrial organoids are limited. In particular, there are not many reports on the specific test process, operation steps, culture conditions, and medium formulations.

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