Ovarian normal and tumor-associated fibroblasts retain in vivo stromal characteristics in a 3-D matrix-dependent manner.
OBJECTIVE
Due to a lack of experimental systems, little is known about ovarian stroma. Here, we introduce an in vivo-like 3-D system of mesenchymal stromal progression during ovarian tumorigenesis to support the study of stroma permissiveness in human ovarian neoplasias.


METHODS
To sort 3-D cultures into 'normal,' 'primed' and 'activated' stromagenic stages, 29 fibroblastic cell lines from 5 ovarian tumor samples (tumor ovarian fibroblasts, TOFs) and 14 cell lines from normal prophylactic oophorectomy samples (normal ovarian fibroblasts, NOFs) were harvested and characterized for their morphological, biochemical and 3-D culture features.


RESULTS
Under 2-D conditions, cells displayed three distinct morphologies: spread, spindle, and intermediate. We found that spread and spindle cells have similar levels of alpha-SMA, a desmoplastic marker, and consistent ratios of pFAKY(397)/totalFAK. In 3-D intermediate cultures, alpha-SMA levels were virtually undetectable while pFAKY(397)/totalFAK ratios were low. In addition, we used confocal microscopy to assess in vivo-like extracellular matrix topography, nuclei morphology and alpha-SMA features in the 3-D cultures. We found that all NOFs presented 'normal' characteristics, while TOFs presented both 'primed' and 'activated' features. Moreover, immunohistochemistry analyses confirmed that the 3-D matrix-dependent characteristics are reminiscent of those observed in in vivo stromal counterparts.


CONCLUSIONS
We conclude that primary human ovarian fibroblasts maintain in vivo-like (staged) stromal characteristics in a 3-D matrix-dependent manner. Therefore, our stromal 3-D system offers a tool that can enhance the understanding of both stromal progression and stroma-induced ovarian tumorigenesis. In the future, this system could also be used to develop ovarian stroma-targeted therapies.
Quiros
Roderick M
RM
Basic Science, Fox Chase Cancer Center, Philadelphia, PA 19111-2497, USA.
Valianou
Matthildi
M
Kwon
Youngjoo
Y
Brown
Kimberly M
KM
Godwin
Andrew K
AK
Cukierman
Edna
E
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D006801:Humans; D051379:Mice; D009367:Neoplasm Staging; D010051:Ovarian Neoplasms / Q000473:pathology* / Q000503:physiopathology*; D010053:Ovary / Q000166:cytology / Q000502:physiology*; D011817:Rabbits; D017154:Stromal Cells / Q000502:physiology*