SYSTEMS AND METHODS FOR CULTURING CELLS IN SUSPENSION

    公开(公告)号:US20220073868A1

    公开(公告)日:2022-03-10

    申请号:US17422490

    申请日:2020-01-10

    Abstract: A method of culturing adherent cells in suspension is provided that includes culturing adherent cells on a first substrate in a first suspension, harvesting the adherent cells from the first substrate, and transfecting the harvested adherent cells using electro-poration. The method also includes, after the step of transfecting, suspending the transfected adherent cells in a second suspension. A dissolution process for dissolving the second microcarrier particle to harvest the cells or cell products is also provided. This dissolution process includes adding a chelator, such as EDTA, to the second suspension for a predetermined time to separate the cells from the second microcarrier; and isolating the cells or cell products from a remainder of the second suspension after the predetermined time. The dissolution process is performed without enzymes such as pectinase or protease.

    HIGH DENSITY 3D HEPATOCYTE SPHEROID PLATFORM FOR DRUG ADME STUDIES

    公开(公告)号:US20210018492A1

    公开(公告)日:2021-01-21

    申请号:US16980189

    申请日:2019-03-12

    Abstract: The present disclosure relates to methods for evaluating the interaction of a candidate compound on 3D hepatocyte spheroid in an invitro culture, including evaluating the metabolism of a candidate compound, for use in various biochemical and molecular biology studies. The methods are performed in labware that combine 3D spheroid culture with micro-patterned design that allows for multiple to several hundreds of spheroids to be treated under the same conditions and to produce sufficient materials (e.g., parent drug, drug metabolites, DNA, RNA, and proteins from cells) and higher detection signal intensity for ADME/Tox (absorption, distribution, metabolism, excretion and toxicity) studies. The methods allow for, among other uses, the investigation and generation of accurate invitro intrinsic clearance data and thus more accurate prediction of in vivo clearance, particularly with low clearance compounds.

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