Special microenvironmental conditions are required to induce and/or maintain specific qualities of differentiated cells. An important parameter is the three-dimensional tissue architecture that cannot be reproduced in conventional monolayer systems. Advanced tissue culture systems will meet many of these demands, but may reach their limits, especially when gradients of specific substances over distinct tissue layers must be established for long-term culture. These limitations may be overcome by incorporating microstructures into tissue-like culture systems. The microstructured cell support presented consists of a flat array of 625 cubic microcontainers with porous bottoms, in which cells can be supplied with specific media from both sides of the tissue layer. Permanent cell lines and primary rat hepatocytes have been used to test the culture system. In order to define reproducible conditions for tissue formation and for cell adherence to the structure, several ECM (extracellular matrix) components were tested for coating of microstructured substrata. The described tissue culture system offers great flexibility in adapting the cell support to specific needs.

1.
Ammann
P.
, and
Maier
P.
,
1997
, “
Preservation and Inducibility of Xenobiotic Metabolism in Long-Term Cultures of Adult Rat Liver Cell Aggregates
,”
Toxicology in Vitro
, Vol.
11
, pp.
43
56
.
2.
Bader
A.
,
Knop
E.
,
Kern
A.
,
Bo¨
K.
,
Fru¨hauf
N.
,
Crome
O.
,
Esselmann
H.
,
Pape
C.
,
Kempka
G.
, and
Sewing
K. Fr.
,
1996
, “
3-D Coculture of Hepatic Sinusoidal Cells With Primary Hepatocytes—Design of an Organotypical Model
,”
Exp. Cell Res.
, Vol.
226
, pp.
223
233
.
3.
Berry
M. N.
, and
Friend
D. S.
,
1969
, “
High-Yield Preparation of Isolated Rat Liver Parenchymal Cells
,”
J. Cell Biol.
, Vol.
43
, pp.
506
520
.
4.
Bhatia
S. N.
,
Yarmush
M. L.
, and
Toner
M.
,
1996
, “
Controlling Cell Interactions by Micropatterning in Co-cultures: Hepatocyte and 3T3 Fibroblasts
,”
J. Biomed. Material Res.
, Vol.
34
, pp.
189
199
.
5.
Dunn
J. C. Y.
,
Tompkins
R. G.
, and
Yarmush
M. L.
,
1992
, “
Hepatocytes in Collagen Sandwich: Evidence for Transcriptional and Translational Regulation
,”
J. Cell Biol
, Vol.
116
, pp.
1043
1053
.
6.
Ezzell
R. M.
,
Toner
M.
,
Hendricks
K.
,
Dunn
J. C-Y.
,
Tompkins
R. G.
, and
Yarmush
M. L.
,
1993
, “
Effect of Collagen Gel Configuration on the Cytoskeleton in Cultured Rat Hepatocytes
,”
Exp. Cell Res.
, Vol.
208
, pp.
442
452
.
7.
Glaser, L., 1982, “Cell Recognition: Phenomena in Search of Molecules,” Cellular Recognition, W. A. Frazier, ed., Alan R. Liss, Inc., New York, pp. 759–767.
8.
Jauregui
H. O.
,
Chowdhury
N. R.
, and
Chowdhury
J. R.
,
1996
, “
Use of Mammalian Liver Cells for Artificial Liver Support
,”
Cell Transplantation
, Vol.
5
, pp.
353
367
.
9.
Kleinmann
H. K.
,
Klebe
R. J.
, and
Martin
G. R.
,
1981
, “
Role of Collagenous Matrices in the Adhesion and Growth of Cells
,”
J. Cell Biol.
, Vol.
88
, pp.
473
485
.
10.
Koebe
H. G.
,
Pahernik
S.
,
Eyer
P.
, and
Schildberg
F. W.
,
1994
, “
Collagen Gel Immobilization: A Useful Cell Culture Technique for Long-Term Metabolic Studies on Human Hepatocytes
,”
Xenobiotica
, Vol.
24
, pp.
95
107
.
11.
Koide
N.
,
Sakaguchi
K.
,
Koide
Y.
,
Asano
K.
,
Kawaguchi
M.
,
Matsushima
H.
,
Takenami
T.
,
Shinji
T.
,
Mori
M.
, and
Tsuji
T.
,
1990
, “
Formation of Multicellular Spheroids Composed of Adult Rat Hepatocytes in Dishes With Positively Charged Surfaces and Under Other Nonadherent Environments
,”
Exp. Cell Res.
, Vol.
186
, pp.
227
235
.
12.
Landry
J.
,
Bernier
D.
,
Ouellet
C.
,
Goyette
R.
, and
Marceau
N.
,
1985
, “
Spheroidal Aggregate Culture of Rat Liver Cells: Histotypic Reorganization, Biomatrix Deposition and Maintenance of Functional Activities
,”
J. Cell Biol.
, Vol.
101
, pp.
914
923
.
13.
Macklis
J. D.
,
Sidman
R. L.
, and
Shine
H. D.
,
1985
, “
Cross-Linked Collagen Surface for Cell Culture That is Stable, Uniform and Optically Superior to Conventional Surfaces
,”
In Vitro Cell Dev. Biol
, Vol.
21
, pp.
189
194
.
14.
Mooney
D.
,
Hansen
L.
,
Vacanti
J.
,
Langer
R.
,
Farmer
S.
, and
Ingber
D.
,
1992
, “
Switching From Differentiation to Growth in Hepatocytes: Control by Extracellular Matrix
,”
J. Cell Physiol.
, Vol.
151
, pp.
497
505
.
15.
Schaller
Th.
,
Bier
W.
,
Linder
G.
, and
Schubert
K.
,
1994
, “
Mechanische Mikrostrukturierung metallischer Oberfla¨achen
,”
F&M Feinwerktechnik Mikrotechnik Messtechnik
, Vol.
102
, pp.
274
278
.
16.
Tong
J. Z.
,
De Lagausie
P.
,
Furlan
V.
,
Cresteil
T.
,
Bernard
O.
, and
Alvarez
F.
,
1992
, “
Long-Term Culture of Adult Rat Hepatocyte Spheroids
,”
Exp. Cell Res.
, Vol.
200
, pp.
326
332
.
17.
Wartenberg
M.
, and
Acker
H.
,
1995
, “
Quantitative Recording of Vitality Patterns in Living Multicellular Spheroids by Confocal Microscopy
,”
Micron
, Vol.
26
, pp.
395
404
.
18.
Weibezahn
K. F.
,
Knedlitschek
G.
,
Dertinger
H.
,
Bier
W.
,
Schaller
Th.
, and
Schubert
K.
,
1995
, “
Reconstruction of Tissue Layers in Mechanically Processed Microstructures
,”
J. Exp. Clin. Cancer Res.
, Vol.
14
, pp.
41
42
.
19.
Weibezahn, K. F., Kneditschek, G., Bier, W., and Schaller, Th., 1994, “Mechanically Processed Microstructures Used to Establish an In Vitro Tissue Model,” Micro System Technologies ’94, Reichl, H., and Heuberger, A., eds., vde-verlag gmbh, Berlin, pp. 873–878.
20.
Wu
F. J.
,
Friend
J. R.
,
Hsiao
C. C.
,
Zilliox
M. J.
,
Ko
W. J.
,
Cerra
F. B.
, and
Hu
W. S.
,
1996
, “
Efficient Assembly of Rat Hepatocyte Spheroids for Tissue Engineering Applications
,”
Biotechnol. Bioeng.
, Vol.
50
, pp.
404
415
.
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