Microarray approach to investigate gene expression in human ovarian tissue after xenografting

ABSTRACT: Cryobanking and transplantation of ovarian tissue is a promising approach to restore fertility in cancer patients. However, ischemic stress following avascular ovarian cortex grafting is known to induce stromal tissue fibrosis and alteration in follicular development. The aim of the study is to analyse the impact of freezing-thawing and grafting procedures on gene expression in human ovarian tissue. Frozen-thawed ovarian tissue from 4 patients was xenografted for 7 days in nude mice and one non-grafted fragment was used as control. Immediately after recovery, grafts were processed for RNA extraction and histological analysis. Their expression profile was screened by whole-genome oligonucleotide array (n=4) and validated by reverse-transcriptase polymerase chain analysis (n=10). 84 of the transcripts were significantly altered after 7 days of grafting including matrix metalloproteinase-9 and -14 and angiogenic factors such as Placental growth factor and C-X-C chemokine receptor type 4 (CXCR4). Major biological processes were related to tissue remodelling including secretory processes, cellular adhesion and response to chemical and hormone stimuli. Angiopoietin signaling, interleukin-8 pathway and peroxisome proliferator-activated receptors activation were shown to be differentially regulated. On day 7, overexpression was confirmed by PCR for interleukin-8, transforming growth factor-beta 1, matrix metalloproteinase-14 and CXCR4 compared to the nongrafted control. In conclusion, new as well as known genes involved in tissue restructuration and angiogenesis were identified after human ovarian tissue transplantation. This will facilitate the development of strategies to optimize grafting techniques. In this study, ovarian tissue was obtained from 4 patients. Each biopsy was divided into 2 fragments. One fragment was frozen/thawed and placed in TRIzol reagent (Invitrogen) for RNA extraction. The other fragment was frozen-thawed and grafted in the intraperitoneal cavity of a nude mice. After 7 days graft were recovered and total RNA was extracted.

ORGANISM(S): Homo sapiens

SUBMITTER: Jérôme Ambroise 

PROVIDER: E-GEOD-47555 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress