Table 1.
Cell-free DNA level in follicular fluid pools according to the patients’ clinical characteristics.
Table 2.
Cell-free DNA level in follicular fluid pools according to controlled ovarian stimulation protocols and ovarian response to stimulation.
Fig 1.
Cell-free DNA level in follicular fluid pools according to the patients’ ovarian reserve status, ovarian reserve parameters and infertility length.
A, Follicular fluid cfDNA content in patients with normal ovarian reserve versus patients with ovarian reserve disorders (ovarian insufficiency and polycystic ovary syndrome); *p = 0.03. B, Follicular fluid cfDNA content according to the ovarian reserve parameters; left panel: AFC (<10 versus ≥ 10, *p = 0.04); right panel: AMH (≤ 1 versus > 1 ng/ml, *p = 0.06). C, Follicular fluid cfDNA levels according to the infertility length (1 versus ≥ 5 years, *p = 0.049).
Fig 2.
CfDNA level in follicular fluid pools according to the ovarian stimulation protocol and ovarian response.
A, Follicular fluid cfDNA content according to the length of ovarian stimulation (≤ 10 versus > 10 days, *p = 0.008). B, Follicular fluid cfDNA content according to the total dose of gonadotropins (<3000 versus ≥3000 IU/l, *p = 0.01). C, Follicular fluid cfDNA content according to the number of retrieved oocytes (≤ 6 versus > 6 oocytes, *p = 0.045).
Fig 3.
CfDNA level in follicular fluid pools according to the embryo outcome at day 2 and 3.
A, Follicular Fluid cfDNA content according to the total number of embryos at day 2 (≤ 2 versus > 2, *p = 0.03). B, Follicular Fluid cfDNA content according to, left panel: the number of top quality (grade 1–2) embryos per patient (0 versus ≥ 1, *p = 0.002) at day 2, right panel: ratio between number of top quality embryos and total number of embryos (< 0.2 versus ≥ 0.2, *p = 0.04) at day 2. C, Follicular Fluid cfDNA content according to, left panel: number of top quality (grade 1–2) embryos per patient (0 versus ≥ 1, *p = 0.006) at day 3, right panel: ratio between number of top quality embryo and total number of embryos (< 0.2 versus ≥ 0.2, *p = 0.02) at day 3. D, Follicular Fluid cfDNA content according to, left panel: fragmentation rate at day 3 (<20% versus ≥ 20%, p = 0.18) and right panel: ratio between blastomere number and total embryo number at day 3 (<6 versus 6–8, *p = 0.02).
Table 3.
Cell-free DNA levels in follicular fluid pools according to the embryo development outcome at early stages (day 2 and day 3).
Fig 4.
ROC curve to evaluate the predictive value of follicular fluid cfDNA level for clinical pregnancy outcome in a multivariate model (including the rank of IVF/ICSI attempts and the number of embryos): area under the curve = 0.73 [0.66–0.87], sensitivity = 60%, specificity = 88%.
Table 4.
Multivariate logistic model showing the prediction of clinical pregnancy according to the cell-free DNA level in follicular fluid pools.
Fig 5.
Schematic model summarizing the significant relationships between cell-free DNA levels in human follicular fluid (FF) and: infertility length, ovarian reserve status, ovarian stimulation, ovarian response to stimulation, embryo development and clinical pregnancy outcomes, respectively.
High cell-free DNA levels in human FF reflect a poor quality of follicular micro-environment and consequently are related to a poor IVF prognosis. r-FSH, recombinant FSH.