Full text
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| Author(s): |
Caroline Ranéa
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD) |
| Defense date: | 2020-11-27 |
| Examining board members: |
Jorge Hallak;
Paula Intasqui Lopes;
Marcilio Nichi
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| Advisor: | Jorge Hallak |
| Abstract | |
The aim of this study was to standardize the ideal sperm incubation period \"in vitro\", and to determine the favorable culture medium for sperm. 82 asthenozoosperm seminal samples were included, which were divided into two distinct protocols [protocol I (n= 28) and protocol II (n= 54)]. In protocol I, fresh samples (without culture medium - T0) were incubated at 36.6ºC in a 5% CO2 atmosphere for 4 periods: T1 (1 hour), T2 (2 hours), T3 (3 hours) and T4 (4 hours). In protocol II, the samples were processed using the batch density gradient method or simple washing and then were incubated for 2 hours (period defined by protocol I) in different commercially available culture media: a) Human Tubal Fluid - HTF® + 15% Synthetic Serum Substitute - SSS®, b) Single Continuous Culture Medium - CSCM® + 10% Human Serum Albumin - HSA®, c) Sperm Rinse medium - SR® + fructose and carnitine, d) In Vitro Fertilization Medium - G- IVF® + fructose + 15% SSS®. Seminal parameters and functional tests [Sperm Computer Analysis (SCA®), Sperm Chromatin Structural Assay (SCSA®); Reactive Oxygen Species level (ROS), sperm mitochondrial activity (DAB)] were analyzed characterizing groups pre and post-incubation. For statistical analysis we used analysis of variance (ANOVA), Pearson\'s correlation coefficient, Student\'s t test and we adopted P < 0.05. In protocol I, we observed a significant increase in the progressive motility of sperm (T2; P <0.003) incubated for 2 hours when compared with T1, T3 and T4. Furthermore, in protocol II we can infer that the CSCM® and G-IVF® culture media are suitable for culturing human sperm cells, since they brought benefits to sperm motility (Progressive Motility (MP) P < 0.001 and Total Motility (MT) P = 0.025 vs. MP P = 0.023) respectively. For the other culture media, we did not see any significant difference. In conclusion, asthenozoospermic seminal samples sent for Assisted Human Reproduction techniques, can be incubated for 2 hours with CSCM® + HSA® culture medium, in order to improve the motility percentage of the sperm. (AU) | |
| FAPESP's process: | 17/03599-1 - Determination and standardization of culture medium favorable for sperm incubation |
| Grantee: | Caroline Ranéa |
| Support Opportunities: | Scholarships in Brazil - Master |