In vitro cultivation system suitable for prolonged cultivation of human granulosa cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F65269705%3A_____%2F19%3A00071863" target="_blank" >RIV/65269705:_____/19:00071863 - isvavai.cz</a>

Výsledek na webu

<a href="http://cost-cellfit.eu/wp-content/uploads/2019/10/Proceedings-Annual-Meeting2019.pdf" target="_blank" >http://cost-cellfit.eu/wp-content/uploads/2019/10/Proceedings-Annual-Meeting2019.pdf</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

In vitro cultivation system suitable for prolonged cultivation of human granulosa cells

Popis výsledku v původním jazyce

During follicular growth, the granulosa cells exert several crucial functions in ovarian physiology: they form a functional syncytium surrounding and nursing the oocyte and produce steroids secreted into the bloodstream to orchestrate various organs of the female body for ovulation, fertilization of an oocyte and implantation of an embryo. For this reason, granulosa cells are crucial for many functions and suitable for experimental testing. However, when the granular cells are cultured as monolayers, they progressively lose most of their characteristics, like the presence of follicle- stimulating hormone receptor. In this experiment, we tested GrowDex as a matrix for three-dimensional (3D) culture system which allowed survival and growth of a population of granular cells isolated after denudation of matured human oocytes. GrowDex is an animal free nanofibrilal cellulose hydrogel which makes fibrils with length of several micrometres and diameter of 4-100 nanometres. Granular cells were cultured in a 4well NUNC dish for 16 days and cells from one woman were divided half for standard 2D cultivation and half for 3D (GrowDex) cultivation. After cultivation, proliferation activity was analysed and selected key proteins (gonadotropin receptors, mesenchymal markers) were detected by Western blot. In our experiments, the granular cells cultivated in the 3D system were characterized by higher proliferation activity and forming of spheroids during the first 2 days of cultivation. Detection of the key proteins was much less affected in the 3D than in the 2D cultivation system. Our first experiments demonstrate that a significant number of granulosa cells collected from mature oocytes is able to maintain their functional characteristics over prolonged time periods, when cultured in 3D-matrix GrowDex.

Název v anglickém jazyce

In vitro cultivation system suitable for prolonged cultivation of human granulosa cells

Popis výsledku anglicky

During follicular growth, the granulosa cells exert several crucial functions in ovarian physiology: they form a functional syncytium surrounding and nursing the oocyte and produce steroids secreted into the bloodstream to orchestrate various organs of the female body for ovulation, fertilization of an oocyte and implantation of an embryo. For this reason, granulosa cells are crucial for many functions and suitable for experimental testing. However, when the granular cells are cultured as monolayers, they progressively lose most of their characteristics, like the presence of follicle- stimulating hormone receptor. In this experiment, we tested GrowDex as a matrix for three-dimensional (3D) culture system which allowed survival and growth of a population of granular cells isolated after denudation of matured human oocytes. GrowDex is an animal free nanofibrilal cellulose hydrogel which makes fibrils with length of several micrometres and diameter of 4-100 nanometres. Granular cells were cultured in a 4well NUNC dish for 16 days and cells from one woman were divided half for standard 2D cultivation and half for 3D (GrowDex) cultivation. After cultivation, proliferation activity was analysed and selected key proteins (gonadotropin receptors, mesenchymal markers) were detected by Western blot. In our experiments, the granular cells cultivated in the 3D system were characterized by higher proliferation activity and forming of spheroids during the first 2 days of cultivation. Detection of the key proteins was much less affected in the 3D than in the 2D cultivation system. Our first experiments demonstrate that a significant number of granulosa cells collected from mature oocytes is able to maintain their functional characteristics over prolonged time periods, when cultured in 3D-matrix GrowDex.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10601 - Cell biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů