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2015 ; 11
(1
): 16-29
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Hepatocyte culture in autologous decellularized spleen matrix
#MMPMID25664568
Gao R
; Wu W
; Xiang J
; Lv Y
; Zheng X
; Chen Q
; Wang H
; Wang B
; Liu Z
; Ma F
Organogenesis
2015[]; 11
(1
): 16-29
PMID25664568
show ga
BACKGROUND AND AIMS: Using decellularized scaffold to reengineer liver tissue is
a promising alternative therapy for end-stage liver diseases. Though the
decellularized human liver matrix is the ideal scaffold for reconstruction of the
liver theoretically, the shortage of liver donors is still an obstacle for
potential clinical application. Therefore, an appropriate alternative scaffold is
needed. In the present study, we used a tissue engineering approach to prepare a
rat decellularized spleen matrix (DSM) and evaluate the effectiveness of this DSM
for primary rat hepatocytes culture. METHODS: Rat decellularized spleen matrix
(DSM) was prepared by perfusion of a series of detergents through spleen
vasculature. DSM was characterized by residual DNA and specific extracellular
matrix distribution. Thereafter, primary rat hepatocytes were cultured in the DSM
in a 3-dimensional dynamic culture system, and liver cell survival and biological
functions were evaluated by comparison with 3-dimensional sandwich culture and
also with cultured in decellularized liver matrix (DLM). RESULTS: Our research
found that DSM did not exhibit any cellular components, but preserved the main
extracellular matrix and the intact vasculature evaluated by DNA detection,
histology, immunohistochemical staining, vessel corrosion cast and upright
metallurgical microscope. Moreover, the method of DSM preparation procedure was
relatively simple with high success rate (100%). After seeding primary
hepatocytes in DSM, the cultured hepatocytes survived inside DSM with albumin
synthesis and urea secretion within 10 d. Additionally, hepatocytes in dynamic
culture medium had better biological functions at day 10 than that in sandwich
culture. Albumin synthesis was 85.67 ± 6.34 ?g/10(7) cell/24h in dynamic culture
in DSM compared to 62.43 ± 4.59 ?g/10(7) cell/24h in sandwich culture (P < 0.01)
and to 87.54 ± 5.25 ?g/10(7) cell/24h in DLM culture (P > 0.05); urea release was
32.14 ± 8.62 ?g/10(7) cell/24h in dynamic culture in DSM compared to 20.47 ± 4.98
?g/10(7) cell/24h in sandwich culture (P < 0.05) and to 37.38 ± 7.29 ?g/10(7)
cell/24h cultured in DLM (P > 0.05). CONCLUSION: The present study demonstrates
that DSM can be prepared successfully using a tissue engineering approach. The
DSM is an appropriate scaffold for primary hepatocytes culture.
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