1、Designation: F3206 17Standard Guide forAssessing Medical Device Cytocompatibility with DeliveredCellular Therapies1This standard is issued under the fixed designation F3206; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year
2、of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.1. Scope1.1 This guide outlines the parameters to consider whendesigning in vitro tests to assess the potential impact of adeli
3、very device on a cellular product being dispensed. Thisguide does not provide specific protocols, but rather suggestswhat should be considered the minimum characterizationnecessary to assess device cytocompatibility. Topics discussedinclude selecting an appropriate cell line(s), cell physiologyparam
4、eters to measure, and relevant test procedure variables.Only cells suspended in liquid and infused through a device areconsidered. Cell therapies paired with scaffolds, suspended inhydrogels, or administered via other methods (e.g., tissuegrafting) are not included in the scope of this document. Thi
5、sdocument does not address physical characterization of deliv-ery devices, such as mechanics, composition, or degradation.1.2 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-
6、priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.1.3 This international standard was developed in accor-dance with internationally recognized principles on standard-ization established in the Decision on Principles for theDevelopment of Inter
7、national Standards, Guides and Recom-mendations issued by the World Trade Organization TechnicalBarriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards:2F813 Practice for Direct Contact Cell Culture Evaluation ofMaterials for Medical DevicesF2394 Guide for Measuring Securement of
8、 Balloon Expand-able Vascular Stent Mounted on Delivery SystemF2739 Guide for Quantifying Cell Viability within Bioma-terial ScaffoldsF2809 Terminology Relating to Medical and Surgical Mate-rials and Devices2.2 ISO Standard:3ISO 10993-5 Biological evaluation of medical devices Part5: Tests for in vi
9、tro cytotoxicity3. Terminology3.1 Definitions:3.1.1 cell line, na generic term that includes primary,stem, and immortalized cells.3.1.2 cytocompatible, adjreferring to the lack of unaccept-able impact on a cellular product from interaction with amedical device used for delivery or interaction with m
10、anufac-turing components. For example, a cytocompatible device doesnot unacceptably impact the cells passing through it as tocompromise the potency of the cell therapy product.3.1.3 immortalized cell, na primary cell that has beentransformed or otherwise altered to provide an extendedreplication cap
11、acity beyond that of the originating primary cell.An immortalized cell may be naturally isolated (e.g., cancercell) or purposely transformed in the laboratory.3.1.4 primary cell, na cell with a finite replication poten-tial that has not been biologically altered to promote extendedsurvival. A primar
12、y cell may be frozen or freshly isolated butthe passage history must be known and display demonstrablesenescence.3.1.5 senescence, nthe property attributable to finite cellcultures; namely, their inability to grow beyond a finite numberof population doublings. F28093.1.6 stem cells, nprogenitor cell
13、s capable of self-replication, proliferation, and differentiation. F28093.1.7 viable cell, na cell capable of sustaining metabolicactivity that is structurally intact with a functioning cellmembrane. F27393.2 Definitions of Terms Specific to This Standard:1This test method is under the jurisdiction
14、of ASTM Committee F04 on Medicaland Surgical Materials and Devices and is the direct responsibility of SubcommitteeF04.43 on Cells and Tissue Engineered Constructs for TEMPs.Current edition approved March 1, 2017. Published June 2017. DOI: 10.1520/F320617.2For referenced ASTM standards, visit the AS
15、TM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Available from International Organization for Standardization (ISO), ISOCentral Secretariat, BIBC II, Ch
16、emin de Blandonnet 8, CP 401, 1214 Vernier,Geneva, Switzerland, http:/www.iso.org.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United StatesThis international standard was developed in accordance with internationally recognized principles on sta
17、ndardization established in the Decision on Principles for theDevelopment of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.13.2.1 ancillary equipment, nequipment to be paired withthe delivery device (e.g., fitti
18、ngs, syringes, etc.) to facilitate invitro testing through which the cells will pass but are not partof the delivery device as used in the clinic.3.2.2 delivery device, na medical device designed todeliver therapeutic cells into the body.4. Significance and Use4.1 This guide is designed to assist me
19、dical device manu-facturers as they develop new devices or qualify existingdevices (e.g., catheters, needles) for delivering clinical celltherapies. Cytocompatibility considers the impact of the deliv-ery device on the cells passing through the device during thedelivery procedure. The biological saf
20、ety of the device (e.g.,the devices cytotoxicity) should be addressed via othermethods, such as ISO 10993-5. It is understood that this guidedoes not address testing of specific cellular products withspecific delivery devices. Such testing may be required byregulatory authorities prior to clinical t
21、rial of cellular productor marketing applications. This guide outlines considerationsto make the product qualification procedures more likely tosucceed and more cost effective.4.2 The key aspects of assessing device cytocompatibilityinclude selecting a test cell line or cell lines and determiningthe
22、 cell physiology parameters that will be measured to makea determination of cytocompatibility. Acceptance criteria fordesignating a device as cytocompatible are not detailed here. Itwill be up to the delivery device end user to determine if theresults of a cytocompatibility assessment are sufficient
23、 toconsider that device cytocompatible. Delivery device lot to lotvariability may impact cytocompatibility, therefore validatedmanufacturing processes should be considered when producingdevices for cytocompatibility assessments.5. Cell Selection5.1 The cellular response to delivery device contact wi
24、llcomprise the readout for the cytocompatibility characteriza-tions considered in this guide. Given this, selection of the testcell line to be used is critical. Selecting a cell line thatrepresents the intended use of the delivery device isencouraged, but not required. A single cell line or a panel
25、oflines, whether stem cells, primary cells, immortalized cells, ora mixture thereof, may be necessary to characterize the device.Regardless of the approach selected, the cell line(s) chosenmust demonstrate sensitivity to one or more evaluation param-eters being used to characterize cytocompatibility
26、. The identityof selected cell lines should be authenticated by appropriatemeans. Be mindful that some cell lines have licensing fees orpatent protection which must be addressed. Also consider theavailable supply and potential issues with obtaining additionalcell stocks which may introduce unaccepta
27、ble variability.Ideally, as the field develops further, a reference cell line orpanel of lines may be established for the purpose of devicecytocompatibility testing.5.2 It is critical that the chosen cells are well characterized.Cell and culture condition variables such as growth rate,handling proto
28、cols, media requirements, culture vessel coating,dissociation methods (if adherent), typical morphologies, sta-bility during passaging, acceptable passage number, and simi-lar parameters should all be well established. Delivered cellsmay be assayed for viability or function as part of thecytocompati
29、bility evaluation. It is vital that the baseline viabil-ity and functionality of the cells is established and tracked overtime in order to detect any adverse device impacts.5.3 Animal cell lines may be a possibility for use incytocompatibility testing; however, human cell data is prefer-able. Animal
30、 cells offer an advantage given that the in vitrotesting can be supplemented by delivering cells directly intoestablished animal models for further characterization;however, that is outside the scope of this guide.6. Test Method Design6.1 Determining the device impact on delivered cells neces-sitate
31、s careful characterization of the chosen cell line(s). Theappropriate cell morphology and harvest density for use in anassay must be established. The potential impact of the disso-ciation method if using adherent cells (e.g., trypsin versusnon-enzymatic dissociation) may also need to be considered.
32、Itis assumed that cells in culture will be used for purposes ofexpediency and throughput, but using freshly thawed cellaliquots to mimic a clinical application is also suitable. If thismethod is selected, the baseline physiology of the cells afterthawing must be established. Consideration should be
33、given tohow the liquid chosen for suspension of the cells, or othersuspension liquids, may interact with the delivery devicematerials (e.g., degrade the materials, extract leachables) andimpact cytocompatibility.6.2 Positive and negative assay controls will be includedwhenever possible. The negative
34、 control will be cells harvestedand assayed having never touched the delivery device. Nega-tive control cells should be prepared as if being passed throughthe device (e.g., at the set density, added to any ancillaryequipment required). The positive control will be cells thatshow diminished viability
35、 and/or function. Producing a posi-tive control can be accomplished by different approaches. Aspecific delivery device known to impair cells by impactingcell viability or function would be an ideal positive control.Alternatively, chemical or mechanical treatments known toimpair cell line viability a
36、nd/or function will suffice to dem-onstrate that the cell line is sensitive enough to report theseaspects of physiology. Processing agents used in device manu-facture or known to be present in the manufacturing environ-ment may make suitable positive controls. If using chemical ormechanical treatmen
37、ts, a dose-response relationship should beevident to demonstrate the sensitivity of the chosen cell toimpairment. Different treatments may be used on separatepositive control samples in the same assay to perturb specificfunctions (i.e., multiple positive controls are acceptable).6.3 A key aspect of
38、testing will be selecting device deliveryflow rates that approximate clinical use rates. At a minimum,delivery should be assessed at both a minimum and a maxi-mum approximated clinical use flow rate. Syringe pumps orsimilar calibrated equipment should be used to ensure deliveryconsistency. Since the
39、 delivery flow rate will determine intra-device forces and the impact on delivered cells, when possible,F3206 172the ancillary equipment required to use the device being tested(e.g., syringes, adapter fittings, needles, etc.) should be keptconstant for all testing. Different device sizes may necessi
40、tatemultiple groups of ancillary equipment. The chosen cell line(s)and intended clinical use of the delivery device may dictate thatadditional delivery rates besides the minimum and maximumshould be part of the assessment.6.4 Cell density in the test sample delivered through thedevice may also be im
41、portant as different cell lines showdensity effects. As with delivery rates, careful characterizationof the chosen cell line during assay development will deter-mine if a range of densities should be part of the protocol or ifa single standard density is sufficient. The volume of liquidsuspension to
42、 deliver should be sufficient to account for anydead space that the device will hold and still yield enoughsample volume for all subsequent characterization. The meth-ods used to determine recovery and viability may dictate anacceptable cell density range to use in order to provideconfidence with th
43、e selected method.6.5 Delivery devices should be evaluated in their finishedform as they would be offered for clinical use (i.e., notmodified to facilitate cytocompatibility testing). If testing arange of devices (e.g., an infusion catheter available in a rangeof French sizes and lengths), it is acc
44、eptable to select thelargest and smallest for each parameter as representative of therange as long as the delivery rates, cell densities, and anyrequired ancillary equipment does not change. Smaller devicesdesigned to hold and deliver minimal volumes may require thedelivered cell suspension from mul
45、tiple devices be pooled toacquire sufficient volume for the cell physiology assays. Theage of delivery devices should be considered when selectingdevices to evaluate so that newly manufactured devices andthose at the end of their shelf life are tested separately.6.6 The time interval from preparatio
46、n of the cell suspen-sion for testing to actually performing the physiology assaysshould be part of the experimental design. Some cell lines mayprovide for a longer working time before adverse effectsmanifest which may impact assay throughput and protocolordering.6.7 Cell contact time with the devic
47、e must be considered.Cells allowed to sit in a device in anticipation of delivery mayclump, lose viability, or undergo other undesired physiologicalchanges. A maximum dwell time at ambient temperaturesshould be established for the test cell line in order to standard-ize this aspect of testing. Exten
48、ded dwell time may leachcompounds from the delivery device depending on the cellsuspension medium used.6.8 Delivery device configuration during clinical use shouldbe considered to account for changes in geometry in flexibledevices that may impact cells. For example, if the test deviceis a catheter,
49、consider whether the most demanding configura-tion (e.g., maximum shear forces, most challenging geometry)for infusion is with the catheter in a straight configuration orwith some degree of tortuosity. Guide F2394 and similardocuments may be instructive when designing methods toapproximate a clinically relevant infusion model.Also considerif the cells will be in the device during deployment as this mayimpact cytocompatibility. If so, simulated deployment prior todelivery should be included in the test method.7. Cell Physiology Assessment Post Device Delivery7.1 A multitu
