TY - JOUR
T1 - Ovarian normal and tumor-associated fibroblasts retain in vivo stromal characteristics in a 3-D matrix-dependent manner
AU - Quiros, Roderick M.
AU - Valianou, Matthildi
AU - Kwon, Youngjoo
AU - Brown, Kimberly M.
AU - Godwin, Andrew K.
AU - Cukierman, Edna
N1 - Funding Information:
We thank Dr. A. Klein-Szanto for assistance in pathological observations of immunohistochemistry assays, Ms. C. M. Slater for her expertise in tissue culture, Mrs. O. Villamar for assistance on immunohistochemistry techniques, Drs. R. Castelló-Cros and G. P. Adams, for critical comments, Ms. R. Roth for technical assistance and Mrs. K. Buchheit for proofreading. This study was assisted by the following Fox Chase Cancer Center facilities: Biosample Repository, Histopathology Facility, Cell Culture, Glass-Washing and the Talbot Research Library. In addition, funds supporting this work were obtained from the American Association of Cancer Research (the department specifically disclaims responsibility for any analyses, interpretations, or conclusions), the WW Smith Charitable Trust, the Ovarian Cancer Research Fund, the Ovarian Cancer SPORE at FCCC (P50 CA083638), grants RO1-CA113451 and CA-06927 from the NCI, as well as an appropriation from the Commonwealth of Pennsylvania.
PY - 2008/7
Y1 - 2008/7
N2 - 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 α-SMA, a desmoplastic marker, and consistent ratios of pFAKY397/totalFAK. In 3-D intermediate cultures, α-SMA levels were virtually undetectable while pFAKY397/totalFAK ratios were low. In addition, we used confocal microscopy to assess in vivo-like extracellular matrix topography, nuclei morphology and α-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.
AB - 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 α-SMA, a desmoplastic marker, and consistent ratios of pFAKY397/totalFAK. In 3-D intermediate cultures, α-SMA levels were virtually undetectable while pFAKY397/totalFAK ratios were low. In addition, we used confocal microscopy to assess in vivo-like extracellular matrix topography, nuclei morphology and α-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.
KW - 3-D culture
KW - Extracellular matrix
KW - Tumor stroma
KW - Tumor-associated fibroblasts
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U2 - 10.1016/j.ygyno.2008.03.006
DO - 10.1016/j.ygyno.2008.03.006
M3 - Article
C2 - 18448156
AN - SCOPUS:45549107567
VL - 110
SP - 99
EP - 109
JO - Gynecologic Oncology
JF - Gynecologic Oncology
SN - 0090-8258
IS - 1
ER -