Non-standardized protein background in IVF media linked to
serum-derived albumin supplementation

J 2025

Non-standardized protein background in IVF media linked to serum-derived albumin supplementation

NEZVEDOVÁ, Markéta; Volodymyr POROKH; Tami BOČKOVÁ; Václav PUSTKA; Drahomíra KYJOVSKÁ et. al.

Základní údaje

Originální název

Non-standardized protein background in IVF media linked to serum-derived albumin supplementation

Autoři

NEZVEDOVÁ, Markéta; Volodymyr POROKH; Tami BOČKOVÁ; Václav PUSTKA ; Drahomíra KYJOVSKÁ; Barbora MAIEROVÁ; Soňa KLOUDOVÁ; Pavel OTEVŘEL a Zuzana HOLUBCOVÁ

Vydání

JOURNAL OF ASSISTED REPRODUCTION AND GENETICS, NEW YORK, SPRINGER/PLENUM PUBLISHERS, 2025, 1058-0468

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30214 Obstetrics and gynaecology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 2.700 v roce 2024

Organizační jednotka

Lékařská fakulta

DOI

https://doi.org/10.1007/s10815-025-03616-0

UT WoS

001570393200001

EID Scopus

2-s2.0-105016190630

Klíčová slova anglicky

IVF media composition; Human serum albumin; Spent culture medium; Embryo culture; Proteomics; Protein biomarkers

Štítky

143180, podil

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 24. 11. 2025 11:38, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

Purpose To explore the protein compositional variability of IVF media and identify sources of undeclared contaminants that interfere with the detection of embryo-derived signals. Methods Untargeted and targeted mass spectrometry techniques were used to analyze protein composition in 85 samples of used and unused monophasic IVF media across 13 production lots from two manufacturers. Samples included spent culture media (SCM) from individual embryo cultures, matched controls, and unused (blank) media. Protein-free base media was supplemented with either serum-derived or recombinant human serum albumin (HSA) to evaluate their impact on protein contamination. Results Proteomic analysis revealed that not only SCM but also unconditioned media contained over 700 undeclared human proteins, many of which are known to be implicated in key cellular pathways. No significant differences were observed between the protein profiles of embryos that reached the blastocyst stage (n = 29) and those arrested at cleavage (n = 24). Instead, protein level variation strongly correlated with media production lot, as shown by targeted analysis of 14 candidate proteins and principal component clustering of 53 SCM samples. Analysis of blank media confirmed substantial lot-to-lot heterogeneity. Supplementation experiments demonstrated that serum-derived HSA introduces undeclared, batch-variable proteins into IVF media, contributing to a non-standardized culture environment and confounding the detection of embryo-derived signals. Conclusion Serum-derived HSA was identified as the primary source of protein contamination in IVF media. This overlooked protein background contributes to variability in clinical culture conditions, undermines the reproducibility of secretome analyses, and complicates the discovery of reliable biomarkers in SCM.

Návaznosti

EH23_015/0008175, projekt VaV

Název: Inovace České infrastruktury pro integrativní strukturní biologii

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

LM2023069, projekt VaV

Název: Výzkumná infrastruktura RECETOX

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná infrastruktura RECETOX

MUNI/A/1738/2024, interní kód MU

Název: Zdroje pro tkáňové inženýrství 15

Investor: Masarykova univerzita, Zdroje pro tkáňové inženýrství 15

NU22-08-00543, projekt VaV

Název: Nové biomarkery pro neinvazivní in vitro diagnostiku v klinické embryologii (Akronym: Analyza IVF médií)

Investor: Ministerstvo zdravotnictví ČR, Nové biomarkery pro neinvazivní in vitro diagnostiku v klinické embryologii, Podprogram 1 - standardní

857560, interní kód MU
(Kód CEP: EF17_043/0009632)

Název: CETOCOEN Excellence (Akronym: CETOCOEN Excellence)

Investor: Evropská unie, CETOCOEN Excellence, Spreading excellence and widening participation

90254, velká výzkumná infrastruktura

Název: e-INFRA CZ II

Citovat

NEZVEDOVÁ, Markéta; Volodymyr POROKH; Tami BOČKOVÁ; Václav PUSTKA; Drahomíra KYJOVSKÁ; Barbora MAIEROVÁ; Soňa KLOUDOVÁ; Pavel OTEVŘEL a Zuzana HOLUBCOVÁ. Non-standardized protein background in IVF media linked to serum-derived albumin supplementation. JOURNAL OF ASSISTED REPRODUCTION AND GENETICS. NEW YORK: SPRINGER/PLENUM PUBLISHERS, 2025, roč. 42, č. 10, s. 3535-3549. ISSN 1058-0468. Dostupné z: https://doi.org/10.1007/s10815-025-03616-0.

@article{2519167,
   author = {Nezvedová, Markéta and Porokh, Volodymyr and Bočková, Tami and Pustka, Václav and Kyjovská, Drahomíra and Maierová, Barbora and Kloudová, Soňa and Otevřel, Pavel and Holubcová, Zuzana},
   article_location = {NEW YORK},
   article_number = {10},
   doi = {https://doi.org/10.1007/s10815-025-03616-0},
   keywords = {IVF media composition; Human serum albumin; Spent culture medium; Embryo culture; Proteomics; Protein biomarkers},
   language = {eng},
   issn = {1058-0468},
   journal = {JOURNAL OF ASSISTED REPRODUCTION AND GENETICS},
   note = {Nedoplňovat návaznost na EF17_043/0009632.},
   title = {Non-standardized protein background in IVF media linked to serum-derived albumin supplementation},
   url = {https://link.springer.com/article/10.1007/s10815-025-03616-0},
   volume = {42},
   year = {2025}
}

TY  - JOUR
ID  - 2519167
AU  - Nezvedová, Markéta - Porokh, Volodymyr - Bočková, Tami - Pustka, Václav - Kyjovská, Drahomíra - Maierová, Barbora - Kloudová, Soňa - Otevřel, Pavel - Holubcová, Zuzana
PY  - 2025
TI  - Non-standardized protein background in IVF media linked to serum-derived albumin supplementation
JF  - JOURNAL OF ASSISTED REPRODUCTION AND GENETICS
VL  - 42
IS  - 10
SP  - 3535-3549
EP  - 3535-3549
PB  - SPRINGER/PLENUM PUBLISHERS
SN  - 10580468
N1  - Nedoplňovat návaznost na EF17_043/0009632.
KW  - IVF media composition
KW  - Human serum albumin
KW  - Spent culture medium
KW  - Embryo culture
KW  - Proteomics
KW  - Protein biomarkers
UR  - https://link.springer.com/article/10.1007/s10815-025-03616-0
N2  - Purpose To explore the protein compositional variability of IVF media and identify sources of undeclared contaminants that interfere with the detection of embryo-derived signals. Methods Untargeted and targeted mass spectrometry techniques were used to analyze protein composition in 85 samples of used and unused monophasic IVF media across 13 production lots from two manufacturers. Samples included spent culture media (SCM) from individual embryo cultures, matched controls, and unused (blank) media. Protein-free base media was supplemented with either serum-derived or recombinant human serum albumin (HSA) to evaluate their impact on protein contamination. Results Proteomic analysis revealed that not only SCM but also unconditioned media contained over 700 undeclared human proteins, many of which are known to be implicated in key cellular pathways. No significant differences were observed between the protein profiles of embryos that reached the blastocyst stage (n = 29) and those arrested at cleavage (n = 24). Instead, protein level variation strongly correlated with media production lot, as shown by targeted analysis of 14 candidate proteins and principal component clustering of 53 SCM samples. Analysis of blank media confirmed substantial lot-to-lot heterogeneity. Supplementation experiments demonstrated that serum-derived HSA introduces undeclared, batch-variable proteins into IVF media, contributing to a non-standardized culture environment and confounding the detection of embryo-derived signals. Conclusion Serum-derived HSA was identified as the primary source of protein contamination in IVF media. This overlooked protein background contributes to variability in clinical culture conditions, undermines the reproducibility of secretome analyses, and complicates the discovery of reliable biomarkers in SCM.
ER  -

NEZVEDOVÁ, Markéta; Volodymyr POROKH; Tami BOČKOVÁ; Václav PUSTKA; Drahomíra KYJOVSKÁ; Barbora MAIEROVÁ; Soňa KLOUDOVÁ; Pavel OTEVŘEL a Zuzana HOLUBCOVÁ. Non-standardized protein background in IVF media linked to serum-derived albumin supplementation. \textit{JOURNAL OF ASSISTED REPRODUCTION AND GENETICS}. NEW YORK: SPRINGER/PLENUM PUBLISHERS, 2025, roč.~42, č.~10, s.~3535-3549. ISSN~1058-0468. Dostupné z: https://doi.org/10.1007/s10815-025-03616-0.

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