title: Basic fibroblast growth factor promotes the development of human ovarian early follicles during growth in vitro.
abstract: OBJECTIVE
  What is the effect of basic fibroblast growth factor (bFGF) on the growth of individual early human follicles in a three-dimensional (3D) culture system in vitro?
  
  
  CONCLUSIONS
  The addition of 200 ng bFGF/ml improves human early follicle growth, survival and viability during growth in vitro.
  
  
  BACKGROUND
  It has been demonstrated that bFGF enhances primordial follicle development in human ovarian tissue culture. However, the growth and survival of individual early follicles in encapsulated 3D culture have not been reported.
  
  
  METHODS
  The maturation in vitro of human ovarian follicles was investigated. Ovarian tissue (n= 11) was obtained from 11 women during laparoscopic surgery for gynecological disease, after obtaining written informed consent. One hundred and fifty-four early follicles were isolated by enzymic digestion and mechanical disruption. They were individually encapsulated into alginate (1% w/v) and randomly assigned to be cultured with 0, 100, 200 or 300 ng bFGF/ml for 8 days.
  
  
  METHODS
  Individual follicles were cultured in minimum essential medium α (αMEM) supplemented with bFGF. Follicle survival and growth were assessed by microscopy. Follicle viability was evaluated under confocal laser scanning microscope following Calcein-AM and Ethidium homodimer-I (Ca-AM/EthD-I) staining.
  
  
  RESULTS
  After 8 days in culture, all 154 follicles had increased in size. The diameter and survival rate of the follicles and the percentage with good viability were significantly higher in the group cultured with 200 ng bFGF/ml than in the group without bFGF (P < 0.05). The percentage of follicles in the pre-antral stage was significantly higher in the 200 ng bFGF/ml group than in the group without bFGF (P < 0.05), while the percentages of primordial and primary follicles were significantly lower (P < 0.05).
  
  
  CONCLUSIONS
  The study focuses on the effect of bFGF on the development of individual human early follicles in 3D culture in vitro and has limited ability to reveal the specific effect of bFGF at each different stage. The findings highlight the need to improve the acquisition and isolation of human ovarian follicles.
  
  
  CONCLUSIONS
  The in vitro 3D culture of human follicles with appropriate dosage of bFGF offers an effective method to investigate their development. Moreover, it allows early follicles to be cultured to an advanced stage and therefore has the potential to become an important source of mature oocytes for assisted reproductive technology; particularly as an option for fertility preservation in women, including patients with cancer.
  
  
  BACKGROUND
  This work was supported by the National Basic Research Program of China (2011|CB944504, 2011CB944503) and the National Natural Science Foundation of China (81200470, 81000275, 31230047, 8110197). There are no conflicts of interest to declare.
authors.affiliation: Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, No. 49 North HuaYuan Road, HaiDian District, Beijing 100191, China.
authors.forename: Tian-ren
authors.initials: TR
authors.lastname: Wang
authors.forename: Li-ying
authors.initials: LY
authors.lastname: Yan
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.forename: Cui-ling
authors.initials: CL
authors.lastname: Lu
authors.forename: Xi
authors.initials: X
authors.lastname: Xia
authors.forename: Tai-lang
authors.initials: TL
authors.lastname: Yin
authors.forename: Xiao-hui
authors.initials: XH
authors.lastname: Zhu
authors.forename: Jiang-man
authors.initials: JM
authors.lastname: Gao
authors.forename: Ting
authors.initials: T
authors.lastname: Ding
authors.forename: Wei-hong
authors.initials: WH
authors.lastname: Hu
authors.forename: Hong-yan
authors.initials: HY
authors.lastname: Guo
authors.forename: Rong
authors.initials: R
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Qiao
mesh_terms: D000328:Adult; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D006080:Ovarian Follicle / Q000187:drug effects* / Q000254:growth & development*; D046509:Tissue Culture Techniques / Q000379:methods
chemical_list: D016222:Fibroblast Growth Factor 2
keywords: basic fibroblast growth factor; human follicle; in vitro 3D culture; in vitro growth