EN ISO 10993-5:2009

SLOVENSKI STANDARD
01-september-2009
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SIST EN ISO 10993-5:2000
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Biological evaluation of medical devices - Part 5: Tests for in vitro cytotoxicity (ISO 10993
-5:2009)
Biologische Beurteilung von Medizinprodukten - Teil 5: Prüfungen auf in vitro-
Zytotoxizität (ISO 10993-5:2009)
Évaluation biologique des dispositifs médicaux - Partie 5: Essais concernant la
cytotoxicité in vitro (ISO 10993-5:2009)
Ta slovenski standard je istoveten z: EN ISO 10993-5:2009
ICS:
11.100.20 %LRORãNRRYUHGQRWHQMH Biological evaluation of
PHGLFLQVNLKSULSRPRþNRY medical devices
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 10993-5
NORME EUROPÉENNE
EUROPÄISCHE NORM
June 2009
ICS 11.100.20 Supersedes EN ISO 10993-5:1999
English Version
Biological evaluation of medical devices - Part 5: Tests for in
vitro cytotoxicity (ISO 10993-5:2009)
Évaluation biologique des dispositifs médicaux - Partie 5: Biologische Beurteilung von Medizinprodukten - Teil 5:
Essais concernant la cytotoxicité in vitro (ISO 10993- Prüfungen auf In-vitro-Zytotoxizität (ISO 10993-5:2009)
5:2009)
This European Standard was approved by CEN on 17 April 2009.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN Management Centre or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as the
official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2009 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 10993-5:2009: E
worldwide for CEN national Members.

Contents Page
Foreword .3
Annex ZA (informative)  Relationship between this International Standard and the Essential
Requirements of EU Directive 93/42/EEC .4
Annex ZB (informative)  Relationship between this International Standard and the Essential
Requirements of EU Directive 90/385/EEC .5

Foreword
This document (EN ISO 10993-5:2009) has been prepared by Technical Committee ISO/TC 194 "Biological
evaluation of medical devices" in collaboration with Technical Committee CEN/TC 206 “Biological evaluation
of medical devices” the secretariat of which is held by NEN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by December 2009, and conflicting national standards shall be withdrawn
at the latest by December 2009.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 10993-5:1999.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association, and supports essential requirements of EC Directives.
For relationship with EC Directives, see informative Annex ZA and ZB, which are integral parts of this
document.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech
Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia,
Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 10993-5:2009 has been approved by CEN as EN ISO 10993-5:2009 without any
modifications.
Annex ZA
(informative)
Relationship between this International Standard and the Essential
Requirements of EU Directive 93/42/EEC
This International Standard has been prepared under a mandate given to CEN by the European Commission
and the European Free Trade Association to provide a means of conforming to Essential Requirements of the
New Approach Directive 93/42/EEC on medical devices.

Once this standard is cited in the Official Journal of the European Communities under that Directive and has
been implemented as a national standard in at least one Member State, compliance with the clauses of this
standard given in Table ZA.1 confers, within the limits of the scope of this standard, a presumption of
conformity with the corresponding Essential Requirements of that Directive and associated EFTA regulations.

Table ZA.1— Correspondence between this International Standard and Directive 93/42/EEC

Clause(s)/sub-clause(s) of Essential Requirements (ERs) of Directive Qualifying
this International Standard 93/42/EEC remarks/Notes

4, 5, 6, 7, 8, 9, 10 Annex I:
7.1, 7.2, 7.5
WARNING: Other requirements and other EU Directives may be applicable to the product(s) falling within the
scope of this International standard.

Annex ZB
(informative)
Relationship between this International Standard and the Essential
Requirements of EU Directive 90/385/EEC
This International Standard has been prepared under a mandate given to CEN by the European Commission
and the European Free Trade Association to provide a means of conforming to Essential Requirements of the
New Approach Directive 90/385/EEC on active implantable medical devices.

Once this standard is cited in the Official Journal of the European Communities under that Directive and has
been implemented as a national standard in at least one Member State, compliance with the clauses of this
standard given in Table ZB.1 confers, within the limits of the scope of this standard, a presumption of
conformity with the corresponding Essential Requirements of that Directive and associated EFTA regulations.
Table ZB.1 — Correspondence between this International Standard and Directive 90/385/EEC

Clause(s)/sub-clause(s) of Essential Requirements (ERs) of Qualifying
this International Standard Directive 90/385/EEC remarks/Notes

4, 5, 6, 7, 8, 9, 10 Annex I:
WARNING — Other requirements and other EU Directives may be applicable to the product(s) falling within
the scope of this standard.
INTERNATIONAL ISO
STANDARD 10993-5
Third edition
2009-06-01
Biological evaluation of medical
devices —
Part 5:
Tests for in vitro cytotoxicity
Évaluation biologique des dispositifs médicaux —
Partie 5: Essais concernant la cytotoxicité in vitro

Reference number
ISO 10993-5:2009(E)
©
ISO 2009
ISO 10993-5:2009(E)
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ISO 10993-5:2009(E)
Contents Page
Foreword. iv
Introduction . vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions. 1
4 Sample and control preparation. 2
4.1 General. 2
4.2 Preparation of liquid extracts of material. 3
4.3 Preparation of material for direct-contact tests .4
4.4 Preparation of controls . 5
5 Cell lines . 5
6 Culture medium. 5
7 Preparation of cell stock culture. 6
8 Test procedures . 6
8.1 Number of replicates . 6
8.2 Test on extracts . 6
8.3 Test by direct contact. 7
8.4 Test by indirect contact. 7
8.5 Determination of cytotoxicity . 9
9 Test report . 10
10 Assessment of results. 11
Annex A (informative) Neutral red uptake (NRU) cytotoxicity test. 12
Annex B (informative) Colony formation cytotoxicity test. 19
Annex C (informative) MTT cytotoxicity test . 24
Annex D (informative) XTT cytotoxicity test. 29
Bibliography . 34

ISO 10993-5:2009(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with t
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