Background
Methods
Materials
Experimental Design
Experiments | N° samples | Parameter evaluated | |
---|---|---|---|
1 |
In vitro: effect of different amounts of PC on embryo development | 705 oocytes | • Rate of embryos • Total cell numbers per blastocyst |
2 |
In vivo: evaluation of endometrial cell proliferation, after in vivo PC administration | 4 RBCs | Microscopic nuclear count of cells expressing Ki-67 |
3 |
In vivo: embryo implantation and development after intrauterine administration of PC | 30 Treated RBCs 30 Control RBCs | Rate of pregnancy |
Animals and repeat breeder syndrome diagnosis
Preparation of Platelet Concentrate (PC)
Experiment 1: effect of PC on in vitro embryo production
Experiment 2: immunohistochemical analysis of endometrium after in vivo PC administration
Experiment 3: in vivo effectiveness of intrauterine administration of PC at 48 h after AI in RBCs
Bacteriological examination of cervical mucus
Intrauterine administration of platelet concentrate
Progesterone assay
Statistical analysis
Results
Platelet collection efficiency and product sterility test
Experiment 1: effect of PC on in vitro embryo production
Medium | N° of blastocysts at 7 days (%) | N° of embryos observed | Total cell numbers per blastocyst |
---|---|---|---|
CTR (10 % FCS) | 84/240 (35.00 ± 1.72)a
| 29 | 104.6 ± 10.5a
|
FCS 5 % + PC 5 % | 99/230 (43.04 ± 1.42)b
| 31 | 123.4 ± 8.3b
|
PC 10 % | 64/235 (27.23 ± 1.39)c
| 34 | 98.7 ± 6.4c
|
Experiment 2: immunohistochemical analysis of endometrium after in vivo PC administration
1 | 2 | 3 | 4 | ||||
---|---|---|---|---|---|---|---|
Uterine horn | Uterine horn | Uterine horn | Uterine horn | ||||
CTR | TRT | CTR | TRT | CTR | TRT | CTR | TRT |
2 ± 0.7a
| 12 ± 0.2b
| 1 ± 0.2a
| 10 ± 0.6b
| 5 ± 2.9a
| 18 ± 2.3b
| 14 ± 3.8a
| 40 ± 4.2b
|
Experiment 3: in vivo effectiveness of intrauterine administration of PC at 48 hrs after AI in RBCs
Progesterone value (ng/ml) | |||||||
---|---|---|---|---|---|---|---|
N°cows | T0 | T4 | T8 | Pregnant (%) | Not Pregnant (%) | Delivery (%) | |
Control | 30 | 0.40 ± 0.21a
| 2.63 ± 0.96a
| 7.75 ± 2.21a
| 10 (33.33)a
| 20 (66.67)a
| 10 (100) |
Treated | 30 | 0.40 ± 0.17a
| 3.12 ± 0.85a
| 8.33 ± 2.14a
| 21 (70.00)b
| 9 (30.00)b
| 21 (100) |