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Morphologic differentiation of colon carcinoma cell lines HT-29 and HT-29KM in rotating-wall vesselsA new low shear stress microcarrier culture system has been developed at NASA's Johnson Space Center that permits three-dimensional tissue culture. Two established human colon adenocarcinoma cell lines, HT-29, an undifferentiated, and HT-29KM, a stable, moderately differentiated subline of HT-29, were grown in new tissue culture bioreactors called Rotating-Wall Vessels (RWVs). RWVs are used in conjunction with multicellular cocultivation to develop a unique in vitro tissue modeling system. Cells were cultivated on Cytodex-3 microcarrier beads, with and without mixed normal human colonic fibroblasts, which served as the mesenchymal layer. Culture of the tumor lines in the absence of fibroblasts produced spheroidlike growth and minimal differentiation. In contrast, when tumor lines were co-cultivated with normal colonic fibroblasts, initial growth was confined to the fibroblast population until the microcarriers were covered. The tumor cells then commenced proliferation at an accelerated rate, organizing themselves into three-dimensional tissue masses that achieved 1.0- to 1.5-cm diameters. The masses displayed glandular structures, apical and internal glandular microvilli, tight intercellular junctions, desmosomes, cellular polarity, sinusoid development, internalized mucin, and structural organization akin to normal colon crypt development. Differentiated samples were subjected to transmission and scanning electron microscopy and histologic analysis, revealing embryoniclike mesenchymal cells lining the areas around the growth matrices. Necrosis was minimal throughout the tissue masses. These data suggest that the RWV affords a new model for investigation and isolation of growth, regulatory, and structural processes within neoplastic and normal tissue.
Document ID
20050008881
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Goodwin, T. J.
(NASA Johnson Space Center Houston, TX United States)
Jessup, J. M.
Wolf, D. A.
Date Acquired
August 22, 2013
Publication Date
January 1, 1992
Publication Information
Publication: In vitro cellular & developmental biology : journal of the Tissue Culture Association
Volume: 28A
Issue: 1
ISSN: 0883-8364
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other

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