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Three‐dimensional culture of buffalo granulosa cells in hanging drop mimics the preovulatory follicle stage
Author(s) -
Yadav Monica,
Agrawal Himanshu,
Pandey Mamta,
Singh Dheer,
Onteru Suneel K.
Publication year - 2018
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.25909
Subject(s) - follicle , andrology , ovarian follicle , biology , ovary , spheroid , drop (telecommunication) , cell culture , granulosa cell , gene expression , microbiology and biotechnology , chemistry , medicine , endocrinology , gene , biochemistry , genetics , telecommunications , computer science
Granulosa cell (GC) culture models mimicking the intrafollicular environment are limited. Such models have a great potential in reproductive toxicity studies. The buffalo, a monovulatory species like humans, could be a better model than polyovulatory rodents. Therefore, we targeted the development and characterization of three‐dimensional (3D) culture systems for buffalo GCs. The GCs from small ovarian follicles (SF) maintained the CYP19 gene expression for 144 hr in a 2D culture system. Hence, GCs from SF were cultured directly in 3D using hanging drop and Poly‐([2‐hydroxyethyl methacrylate]) (polyHEMA) methods in the DMEM media containing 1 ng/ml FSH and 10 ng/ml IGF‐1 for 144 hr. The expression profile of nine GC‐specific transcripts; CYP19, TNFAIP6, AMH, PTI, NR4A1 , FSHR, RUNX, LHR , and COX2/PTGS2; revealed that 3D‐spheroids developed in hanging drop method maintained the GC phenotype of preovulatory follicles. Therefore, hanging drop method is a best method for culturing GCs to mimic the intrafollicular environment.