ASTM WK36232

    New Test Method for Use of a Centrifugation Method to Quantify/Study Cell-Material Adhesive Interactions

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    Developed by Subcommittee: F04.43 | Committee F04 | Contact Staff Manager



    WK36232

    1. Scope

    1.1 This test method describes a centrifugation cell adhesion assay which can be used to study the alterations in cell adhesion characteristics of chondrocytes cultured on coated or uncoated tissue culture-treated polystyrene as monolayers as a function of cell passage. This test method does not cover methods to verify uniformity of coating for polystyrene surfaces, nor does it cover methods for characterizing surfaces. 1.2 This test method does not cover methods for isolating chondrocytes from the knee joints. 1.3 Include in this section the system of units to be used. Refer to the above ASTM Standards Units toolbar button for a dropdown menu of ASTMs Form and Style Manual statements. 1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

    This test method describes a cell adhesion method that can be used to detach cells (chondrocytes) that have adhered to a tissue culture-treated polystyrene substrate. Propagation of chondrocytes as monolayer cultures on polystyrene surfaces is used extensively for amplifying cell numbers. However, chondrocytes undergo a phenotypic shift when propagated in this manner and display characteristics of more adherent fibroblastic cells. Maintaining the chondrocyte phenotype (Col II production, low adhesion, aggrecan production) is fundamental to the development of safe and effective cartilage repair products. The changes in the adhesion properties of the chondrocytes as the chondrocytes de-differentiate in passage as monolayer cultures include changes in morphology, changes in gene expression of Collagen Type I (Col I) and Collagen type II (Col II), as measured by qRT-PCR and, by changes in biomarkers, as identified by immunostaining,. The information generated by this test method can be used to obtain a semi-quantitative measurement of the adhesion of chondrocytes to an uncoated or pre-coated tissue culture-treated polystyrene surface (i.e., fibronectin, etc.).


    Keywords

    Centrifugation; Cell adhesion assay; Chondrocytes; Serial passage; Monolayer culture; tissue culture-treated polystyrene::

    The title and scope are in draft form and are under development within this ASTM Committee.

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    Work Item Status

    Date Initiated:
    01-30-2012

    Technical Contact:
    David Kaplan

    Item:
    010

    Ballot:
    F04 (14-02)

    Status:
    Ballot Item Approved as F3088-2014 and Pending Publication