Thyroid method 8c: In vitro human adipose stromal cell—human umbilical vein endothelial cell (hASC-HUVEC) vasculogenesis/angiogenesis method

Topic: Endocrine disruption

Test Method Number:

TM2019-19 (EU)

Short Name of TM:

VAAN-hadum

Year received:

2019

Responsible Organisation:

EURL ECVAM - European Union

General Comments:

Method Description

Thyroid hormone disruption can interfere with vasculogenesis and angiogenesis and can be assessed using human primary adipose stromal cells and human primary umbilical vein endothelial cells coculture test without any artificial matrixes. In the method the effect of test item on the formation of vasculature is measured six days after the exposure by quantifying the formed tubules using image analysis. Cell viability test (WST-1) is used to measure general cytotoxicity to clarify if the inhibitory mechanism is related to cytotoxicity.

This method was selected with other methods to assess modulating effects on the thyroid system (OECD, 2014). Information on these selected methods is available in TSAR as ‘Thyroid methods’.

Track Approval Status

Submission
Validation
Peer-review
Recommendation
Regulatory acceptance/Standards

Submission

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Date	Stage	Comment	Documents/Links
Sep 2019	Assessment finalised	Selection of the test method for the Thyroid Validation Study based on the OECD scoping document on 'in vitro and ex vivo methods for the identification of modulators of thyroid hormone signalling' and guidance from the Endocrine Disrupters Testing and Assessment Advisory Group considering scientific advances since the issuing of the OECD scoping document, human relevance, possibility for High Throughput screening and availability of method elements (such as reagents and detection method) to a wide range of future users.	OECD scoping document (OECD, 2014)OECD detailed review paper (OECD, 2012)OECD Endocrine Disrupters Testing and Assessment Advisory Group

Validation

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Date	Stage	Comment	Documents/Links
Nov 2020	Ongoing	Preparatory work carried out by EURL ECVAM before the start of the validation study. During this phase, intellectual property was addressed, legal agreements with method developer, test system supplier and EU-NETVAL laboratory were established, the test system was authenticated, screened for contaminants and information on genetic modification was gathered and adequate training of the EU-NETVAL laboratory was ensured. An outline protocol describing the current status of the method, the quality controlled test system and reference and control items were made available to EU-NETVAL. OECD Good In Vitro Method Practices (GIVIMP, OECD, 2018) has been used as guidance.	OECD Good In Vitro Method Practices (GIVIMP, OECD, 2018)
Feb 2021	Finalised	Final selection of the validation set chemicals to be used in the thyroid validation study PART 2. Selection was based on the advice provided by 15 experts in in vitro, in vivo and clinical thyroid disruption during an expert meeting in November 2019.
Aug 2021	Finalised	Start of the EURL ECVAM coordinated validation study. Method developer 'FICAM (Finland)' and EU-NETVAL facility 'FHAIVE (Finland)' agree to assess the method.
Aug 2022	Finalised	Completion of method definition and description prior to experimental assessment.
Dec 2022	Finalised	PART 1: Assessment of method robustness, reliability and within laboratory reproducibility using few test items.
Feb 2023	Stopped	PART 2: Assessment of the method's mechanistic relevance using selected validation set chemicals covering the underlying mechanism of this method was not performed. Method could be further considered for future validation activities.

Peer-review

Recommendation

Regulatory acceptance/Standards