Hanging drop cultures of human testis and testis cancer samples: a model used to investigate activin treatment effects in a preserved niche.
BACKGROUND
Testicular germ cell tumours of young adults, seminoma or non-seminomas, are preceded by a pre-invasive precursor, carcinoma in situ (CIS), understood to arise through differentiation arrest of embryonic germ cells. Knowledge about the malignant transformation of germ cells is currently limited by the lack of experimental models. The aim of this study was to establish an experimental tissue culture model to maintain normal and malignant germ cells within their niche and allow investigation of treatment effects.


METHODS
Human testis and testis cancer specimens from orchidectomies were cultured in 'hanging drops' and effects of activin A and follistatin treatment were investigated in seminoma cultures.


RESULTS
Testis fragments with normal spermatogenesis or CIS cells were cultured for 14 days with sustained proliferation of germ cells and CIS cells and without increased apoptosis. Seminoma cultures survived 7 days, with proliferating cells detectable during the first 5 days. Activin A treatment significantly reduced KIT transcript and protein levels in seminoma cultures, thereby demonstrating a specific treatment response.


CONCLUSIONS
Hanging drop cultures of human testis and testis cancer samples can be employed to delineate mechanisms governing growth of normal, CIS and tumorigenic germ cells retained within their niche.
University Department of Growth and Reproduction, Rigshospitalet, Blegdamsvej 9, 2100 Copenhagen, Denmark.
A
A
Jørgensen
1] Department of Anatomy and Developmental Biology, Monash University, Melbourne, Clayton, Victoria 3800, Australia [2] Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Clayton, Victoria 3800, Australia.
J
J
Young
University Department of Growth and Reproduction, Rigshospitalet, Blegdamsvej 9, 2100 Copenhagen, Denmark.
J E
JE
Nielsen
Department of Urology, Rigshospitalet, Blegdamsvej 9, 2100, Copenhagen, Denmark.
U N
UN
Joensen
Department of Pathology, Rigshospitalet, Blegdamsvej 9, 2100, Copenhagen, Denmark.
B G
BG
Toft
University Department of Growth and Reproduction, Rigshospitalet, Blegdamsvej 9, 2100 Copenhagen, Denmark.
E
E
Rajpert-De Meyts
1] Department of Anatomy and Developmental Biology, Monash University, Melbourne, Clayton, Victoria 3800, Australia [2] Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Clayton, Victoria 3800, Australia.
K L
KL
Loveland
D028341:Activins / Q000494:pharmacology*; D000328:Adult; D000951:Antigens, Neoplasm / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects; D002278:Carcinoma in Situ / Q000473:pathology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004261:DNA Replication / Q000187:drug effects; D038681:Follistatin / Q000494:pharmacology*; D020869:Gene Expression Profiling; D006801:Humans; D019394:Ki-67 Antigen / Q000032:analysis; D008297:Male; D009024:Morphogenesis / Q000187:drug effects; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D061251:Primary Cell Culture / Q000379:methods; D019009:Proto-Oncogene Proteins c-kit / Q000096:biosynthesis / Q000235:genetics; D018239:Seminoma / Q000473:pathology*; D013091:Spermatogenesis / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects; D013736:Testicular Neoplasms / Q000473:pathology*; D013737:Testis / Q000166:cytology*; D050656:Transcription Factor AP-2 / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D038681:Follistatin; D019394:Ki-67 Antigen; D009363:Neoplasm Proteins; C493679:TFAP2C protein, human; D050656:Transcription Factor AP-2; C422405:activin A; D028341:Activins; D019009:Proto-Oncogene Proteins c-kit