Background
Methods
Determination of APOB concentrations in blood-free FF samples, obtained from 61 female patients
Patients' characteristics
IVF/ICSI procedure
APOB quantification by ELISA in individual FF samples
Quartile model
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Q1: APOB <112 ng/ml
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Q2: 112 <APOB<230 ng/ml
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Q3: 230 <APOB<330 ng/ml
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Q4: APOB>330 ng/ml
Are apolipoprotein B gene expression and protein level regulated in vitro by gonadotropins?
CG isolation
Gonadotropin stimulation of GC primary culture
Quantitative (real-time) PCR: APOB gene expression in GC
Western blot analysis: APOB protein expression in GC
Slot Blot analysis: APOB content of the culture medium
Statistical analyses
Ethical approval
Results
APOB is present in human follicular fluid and provides a new biomarker to predict oocyte and embryo quality
Relationships between APOB concentrations in FF and patient’s characteristics (age, BMI, number of attempts)
Variable | β | Robust SE |
p-value
|
---|---|---|---|
Age (years) | |||
Age >36 versus ≤ 36 | –93.6 | 37.7 | 0.02 |
BMI (kg/m2) 18.5 ≤ BMI < 25 (reference) | − | − | − |
BMI < 18.5 | –162.2 | 48.0 | 0.001 |
25 ≤ BMI < 30 | –59.0 | 72.7 | NS |
BMI ≥ 30 | –142.3 | 48.6 | 0.005 |
Number of attempts | |||
3 and 4 versus 1 and 2 | –148.8 | 51.3 | 0.005 |
APOB concentrations in individual FF samples according to IVF/ICSI outcomes
FF related to | n / number of FF analyzed (%) | APOB (ng/ml) |
p-value
| |
---|---|---|---|---|
Mean ± SD | [95 % CI] | |||
Oocyte | 143 / 201 (71.1) | 271.0 ± 181.8 | [240.9; 301.0] |
p = 0.001 |
Empty zona pellucida (no oocyte) | 58 / 201 (28.9) | 179.0 ± 181.3 | [131.8; 227.2] | |
Normal fertilized oocytes | 107 / 201 (53.2) | 279.8 ± 168 | [247.6; 312.0] |
p = 0.004 |
No fertilized oocytes | 23 / 201 (11.4) | 162.6 ± 192.5 | [79.4; 245.8] | |
Embryo | 92 / 201 (45.8) | 294.3 ± 159.8 | [261.2; 327.4] |
p < 0.001 |
No embryo | 109 / 201(54.2) | 202.6 ± 196.5 | [165.3; 239.9] | |
Early cleavage | 22 / 92 (23.9) | 352.1 ± 192.6 | [266.7; 437.5] | NS |
No early cleavage | 70 / 92 (76.1) | 276.1 ± 144.9 | [241.6; 310.7] | |
Top quality embryo | 45 / 92 (48.9) | 322.1 ± 205.2 | [260.4; 383.7] | NS |
No top quality embryo | 47 / 92 (51.1) | 267.7 ± 93.6 | [240.2; 295.2] |
APOB quartiles (ng/ml) | n / number of FF analyzed (%) |
P
| Crude OR [95 % CI] |
p-value
| Adjusted OR [95 % CI]a
|
p-value
|
---|---|---|---|---|---|---|
Probabilities of obtaining an oocyte | ||||||
Q1 (<112) | 24/50 (48.0) | 0.001 | -- (reference) | -- (reference) | ||
Q2 (≥112, <230) | 40/51 (78.4) | 3.9 [1.65–9.4] | 0.002 | 3.4 [1.04–11.2] | 0.044 | |
Q3 (≥230, <330) | 39/49 (79.6) | 4.2 [1.73–10,3] | 0.002 | 3.3 [1.19–9.08] | 0.022 | |
Q4 (≥330) | 40/51 (78.4) | 3.9 [1.65–9.4] | 0.002 | 3.4 [1.01–11.44] | 0.049 | |
Probabilities of obtaining a fertilized oocyte | ||||||
Q1(<112) | 12/50 (24.0) | <0.001 | -- (reference) | -- (reference) | ||
Q2 (≥112, <230) | 34/51 (66.7) | 6.2 [1.79–21.76] | 0.004 | 7.3 [1.59;33.43] | 0.011 | |
Q3 (≥230, <330) | 33/49 (67.3) | 10.1 [2.38–42.69] | 0.002 | 11.2 [1.67;75.63] | 0.013 | |
Q4 (≥330) | 28/51 (55.0) | 6.4 [1.69–24.37] | 0.006 | 6.3 [1.13;34.86] | 0.035 | |
Probabilities of obtaining an embryo on day 2 | ||||||
Q1(<112) | 5/50 (10.0) | <0.001 | -- (reference) | -- (reference) | ||
Q2 (≥112, <230) | 32/51 (62.8) | 15.2 [5.11–44.95] | <0.001 | 17.3 [2.2;137.1] | 0.007 | |
Q3 (≥230, <330) | 31/49 (63.3) | 15.5 [5.19–46.29] | <0.001 | 15.5 [1.9;130.3] | 0.011 | |
Q4 (≥330) | 24/51 (47.1) | 8 [2.72–23.51] | <0.001 | 8.3 [0.8;82.0] | 0.071 | |
Probabilities of obtaining a top quality embryo on day 2 | ||||||
Q1(<112) | 4/50 (8.0) | 0.047 | -- (reference) | -- (reference) | ||
Q2 (≥112, <230) | 14/51 (27.5) | 4.4 [1.32–14.38] | 0.016 | 4.8 [1.04;22.41] | 0.045 | |
Q3 (≥230, <330) | 13/49 (26.5) | 4.2 [1.24–13.86] | 0.021 | 4.1 [0.75;22.64] | NS | |
Q4 (≥330) | 14/51 (27.4) | 4.4 [1.32–14.38] | 0.016 | 3.9 [0.64;23.91] | NS |