Culture matrix configuration and composition in the maintenance of hepatocyte polarity and function.
Several extracellular matrix (ECM) configurations involving type I collagen and Matrigel were examined for their ability to support differentiated function and polarity of cultured adult rat hepatocytes. Collagen sandwich- and Matrigel-based cultures yielded superior and comparable albumin secretion for at least 2 weeks. In collagen sandwich, hepatocytes were polygonal, and formed multicellular arrays. Collagen sandwich was also found to promote in vivo-like polarization of F-actin, cell adhesion molecules (E-cadherin), and lateral (Na+, K(+)-ATPase, glucose transporter) and apical (dipeptidyl peptidase, aminopeptidase) membrane polarity markers, but not the expression of the gap junction protein connexin 32 and the epidermal growth factor (EGF) receptor. In contrast, hepatocytes cultured in or on Matrigel were more rounded and formed aggregates. Matrigel-based cultures also elicited detectable levels of connexin and EGF receptor and an altered distribution of F-actin, E-cadherin, and apical and lateral membrane proteins. Composite sandwich configurations containing collagen I and Matrigel restored markers lacking in the collagen sandwich, and showed a variable morphology and membrane polarity. Hepatocyte polarity could thus be manipulated by the overall ECM composition. Furthermore, in composite sandwich cultures, these manipulations can be effected largely independent of changes in hepatocyte morphology and albumin secretion.
Moghe
P V
PV
Surgical Services, Massachusetts General Hospital, Boston 02114, USA.
Berthiaume
F
F
Ezzell
R M
RM
Toner
M
M
Tompkins
R G
RG
Yarmush
M L
ML
D000199:Actins / Q000032:analysis; D000818:Animals; D001672:Biocompatible Materials; D015415:Biomarkers; D015820:Cadherins / Q000032:analysis; D002462:Cell Membrane / Q000648:ultrastructure; D002478:Cells, Cultured; D003094:Collagen; D017630:Connexins / Q000032:analysis; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods; D018819:Dipeptidyl Peptidase 4 / Q000032:analysis; D004338:Drug Combinations; D005109:Extracellular Matrix; D005260:Female; D007797:Laminin; D008099:Liver / Q000166:cytology* / Q000502:physiology*; D018613:Microscopy, Confocal; D011509:Proteoglycans; D051381:Rats; D011917:Rats, Inbred Lew; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis; D000097002:Gap Junction beta-1 Protein
D000199:Actins; D001672:Biocompatible Materials; D015415:Biomarkers; D015820:Cadherins; D003094:Collagen; D017630:Connexins; D018819:Dipeptidyl Peptidase 4; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D000254:Sodium-Potassium-Exchanging ATPase; D000097002:Gap Junction beta-1 Protein; C059485:matrigel