title: Effect of KnockOut serum replacement on germ cell development of immature testis tissue culture.
abstract: To compare KnockOut serum replacement (KSR) and fetal bovine serum (FBS) for the development of germ cells. Testicular tissues from Sprague-Dawley rats were cultured for 4 weeks in culture media supplemented with FBS or KSR. Tissue area was measured at the beginning and end of the culturing period. Testicular histology, development of the germ cells, and the diameter of seminiferous tubules were analyzed by hematoxylin and eosin staining. After 4 weeks in culture, apoptosis and expression of the stage-specific spermatogenesis marker genes Kit, Sycp3, and Crisp1 were assayed. Tissues cultured in KSR-supplemented media were able to sustain growth and gradually increase seminiferous tubule diameter during the culture period. In addition, spermatogonia, primary spermatocytes, secondary spermatocytes, and round spermatids were observed after 4 weeks in culture, and reverse transcription-PCR confirmed expression of the marker genes. In comparison, tissues cultured in FBS-supplemented media showed dwindling testicular organization, necrotic seminiferous tubules, and expression of Kit, but inconsistent expression of Sycp3 and Crisp1 KnockOut serum replacement outperforms FBS as a growth media supplements for culturing immature spermatogonial tissue culture.
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
authors.forename: Chunhong
authors.initials: C
authors.lastname: Cai
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Lin
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
authors.forename: Guanghui
authors.initials: G
authors.lastname: Wei
authors.affiliation: Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China. Electronic address: dr_daweihe@163.com.
authors.forename: Dawei
authors.initials: D
authors.lastname: He
mesh_terms: D000818:Animals; D017209:Apoptosis; D002417:Cattle; D003470:Culture Media; D005312:Fetal Blood; D015870:Gene Expression; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012671:Seminiferous Tubules / Q000033:anatomy & histology; D044967:Serum; D013087:Spermatids / Q000166:cytology; D013090:Spermatocytes / Q000166:cytology; D013091:Spermatogenesis / Q000235:genetics; D013093:Spermatogonia / Q000166:cytology; D013094:Spermatozoa / Q000254:growth & development*; D013737:Testis / Q000166:cytology* / Q000254:growth & development; D046509:Tissue Culture Techniques
chemical_list: D003470:Culture Media
keywords: Culture medium; Rat; Serum replacement; Spermatogenesis; Testis; Tissue culture