Evaluating the Efficacy of Exosome-Enriched Culture Media to Enhance In Vitro Maturation and Embryo Development in Poor Ovarian Responders Undergoing IVF: A Randomized Controlled Trial
Phase: NA
Conditions: Poor Ovarian Response, Female Infertility of Other Origin, Oocyte Immaturity
Sponsor: Fertiaguerrevere

Official Title: Exosome-Enriched Culture Media to Enhance In 
Vitro Maturation and Embryo Development in Poor Ovarian 
Responders Undergoing IVF 
NCT Number:  
Document Type: Study Protocol 
Document Date: July 2025 
 
Table of Contents 
1. Background and Rationale 
2. Objectives 
3. Study Design 
4. Study Population 
5. Interventions 
6. Outcome Measures 
7. Statistical Considerations 
8. Ethical Considerations 
9. Data Handling and Confidentiality 
10. 
References 
 
1. Background and Rationale Poor ovarian responders often yield a 
limited number of mature oocytes following ovarian stimulation, resulting 
in reduced chances for successful IVF. Preliminary evidence suggests that 
exosomes, extracellular vesicles rich in bioactive molecules, may support 
oocyte maturation by enhancing intercellular communication in the 
follicular environment. This study aims to investigate whether 
incorporating exosomes into IVF culture media improves the short-term in 
vitro maturation of MI oocytes in poor responder patients. 
2. Objectives 
• Primary Objective: To determine if exosome-enriched culture media 
increases the proportion of MI oocytes maturing to MII within 4 
hours compared to standard IVF media. 

• Secondary Objectives: To assess subsequent fertilization rates, 
embryo quality, blastocyst development, implantation, and clinical 
pregnancy rates. 
3. Study Design A prospective, single-center, randomized controlled trial. 
Patients will undergo standard ovarian stimulation protocols. Retrieved MI 
oocytes will be allocated within each patient to two different culture 
conditions: exosome-enriched media vs. standard media. 
4. Study Population 
• Inclusion Criteria: 
o Women aged 35–42 years 
o Undergoing IVF with antagonist protocols 
o Meet at least two Bologna criteria for poor ovarian response 
o Able to provide written informed consent 
• Exclusion Criteria: 
o Severe male factor infertility requiring surgical sperm retrieval 
o Known untreated hydrosalpinx 
o Uterine anomalies impairing implantation 
o Known chromosomal abnormalities 
o Participation in another investigational study within the last 3 
months 
5. Interventions 
• Control: MI oocytes cultured in pre-equilibrated standard IVF media. 
• Intervention: MI oocytes cultured in the same media supplemented 
with 10 µg/well of GMP-certified exosomes. Maturation status will be 
assessed at 2, 3, and 4 hours. 
6. Outcome Measures 
• Primary Outcome: Proportion of MI oocytes achieving MII 
maturation within 4 hours. 
• Secondary Outcomes: Fertilization rates, day 3 embryo quality, day 
5 blastocyst quality, implantation rate, and clinical pregnancy rate. 
7. Statistical Considerations Categorical outcomes will be compared 
using chi-square or Fisher’s exact tests. Logistic regression will adjust for 

patient age, AMH, and gonadotropin dose. A p-value <0.05 will be 
considered statistically significant. 
8. Ethical Considerations The study will adhere to the Declaration of 
Helsinki and Good Clinical Practice guidelines. Ethical approval will be 
obtained prior to enrollment. All participants will provide written informed 
consent. Confidentiality will be maintained, with data anonymized using 
unique study codes. 
9. Data Handling and Confidentiality Data will be stored in secure, 
password-protected systems. No personally identifiable information will be 
shared or published. 
10. References 
• Melo P, Navarro C, Jones C, Coward K, Coleman L. J Assist Reprod 
Genet. 2020;37(4):855–863. 
• Zhang H, Luo W, Zhu X, et al. Reprod Sci. 2024; Epub ahead of 
print. 
• Additional relevant literature on exosomes and oocyte maturation. 
 
End of Protocol Document. 
 
 
 

Official Title: Evaluating the Efficacy of Exosome-Enriched Culture Media to 
Enhance In Vitro Maturation and Embryo Development in Poor Ovarian Responders 
Undergoing IVF: A Randomized Controlled Trial 
NCT Number: J-298384255-2 
Document Type: Statistical Analysis Plan (SAP) 
Document Date: July 2025 
 
Table of Contents 
1. Introduction 
2. Study Objectives and Endpoints 
3. Sample Size and Power 
4. Analysis Populations 
5. Statistical Methods 
6. Handling of Missing Data 
7. Interim Analyses and Data Monitoring 
8. Reporting Conventions 
 
1. Introduction This Statistical Analysis Plan (SAP) details the planned analyses for 
the study evaluating whether exosome-enriched culture media enhances the in vitro 
maturation of MI oocytes and subsequent embryo development in poor responder 
patients undergoing IVF. 
2. Study Objectives and Endpoints 
• Primary Endpoint: 
o Proportion of MI oocytes maturing to MII within 4 hours of culture. 
• Secondary Endpoints: 
o Fertilization rate (percentage of matured MII oocytes achieving normal 
2PN post-ICSI) 
o Day 3 and blastocyst embryo quality 
o Implantation rate (number of gestational sacs per embryo transferred) 
o Clinical pregnancy rate (confirmed fetal heartbeat at 8–10 weeks) 
3. Sample Size and Power A total of 60 patients will be enrolled, based on 
preliminary estimates indicating that this sample size provides >80% power to detect a 
25% difference in maturation rates at a two-sided alpha of 0.05, assuming maturation 
rates improve from ~40% (control) to ~65% (exosome group). 
4. Analysis Populations 

• Full Analysis Set (FAS): All retrieved MI oocytes allocated to either exosome 
or standard culture conditions. 
• Per Protocol Set (PPS): Oocytes that completed the planned culture period 
without protocol deviations. 
5. Statistical Methods 
• Primary Analysis: 
o Compare the proportion of MI oocytes maturing to MII between exosome 
and standard media using chi-square or Fisher’s exact tests. 
• Secondary Analyses: 
o Compare fertilization rates and embryo quality using similar tests. 
o Implantation and clinical pregnancy rates summarized descriptively and 
analyzed if numbers permit. 
• Adjustment: Logistic regression models may adjust for patient age, baseline 
AMH, and total gonadotropin dose. 
• Significance: Two-sided tests, p<0.05 considered statistically significant. 
6. Handling of Missing Data Missing maturation outcomes will be treated 
conservatively as non-matured. Sensitivity analyses may exclude such oocytes. No 
imputation planned for secondary reproductive outcomes. 
7. Interim Analyses and Data Monitoring No formal interim analyses are planned. 
Data will be reviewed periodically for safety and protocol compliance by the study 
team. 
8. Reporting Conventions Descriptive statistics will include means, medians, 
standard deviations, and ranges. Tables and figures will illustrate maturation rates and 
embryo development outcomes by intervention. 
 
End of Statistical Analysis Plan. 
 
 
 

Official Title: Evaluating the Efficacy of Exosome-Enriched Culture 
Media to Enhance In Vitro Maturation and Embryo Development in Poor 
Ovarian Responders Undergoing IVF: A Randomized Controlled Trial 
NCT Number: J-298384255-2 
Document Type: Informed Consent Form 
Document Date: July 2025 
 
Informed Consent Form (ICF) 
Introduction: You are being invited to participate in a research study 
conducted at [Clinic Name]. This study is investigating whether adding 
exosomes to the culture media used in IVF can improve the maturation of 
certain types of eggs (oocytes) retrieved during your IVF treatment. 
Purpose of the Study: The goal is to determine if supplementing the 
standard IVF media with exosomes can help more of your retrieved eggs 
mature, potentially increasing the number available for fertilization and 
embryo development. 
Procedures: 
• You will undergo your planned ovarian stimulation and egg retrieval 
as part of standard IVF care. 
• After retrieval, some of your immature eggs (MI oocytes) will be 
cultured in either the usual IVF media or IVF media supplemented 
with exosomes. 
• This does not change your medical treatment, medications, or 
procedures. 
Risks and Discomforts: 
• There are no known additional risks to you since the study procedures 
are performed entirely on eggs after retrieval. 
• Standard risks of IVF treatment still apply. 
Benefits: 

• There may be no direct benefit to you, but information gained may 
help improve future IVF treatments. 
Confidentiality: 
• Your data will be assigned a study ID number. No personal identifiers 
will be used in publications or shared outside the study team. 
Voluntary Participation: 
• Your participation is voluntary. Choosing not to participate or to 
withdraw at any time will not affect your medical care. 
Questions: If you have questions about the study, please contact [Study 
Coordinator Name and Contact Info]. 
Consent Statement: By signing below, you confirm that the study has 
been explained to you, your questions have been answered, and you agree 
to participate. 
Participant Name: _______________________________ 
Participant Signature: ___________________________ Date: __________ 
Investigator Signature: __________________________ Date: __________ 
 
End of Informed Consent Form.