Autologous versus pooled human serum for articular chondrocyte growth
Purpose. To evaluate the effect of autologous human serum (AHS) versus pooled human serum (PHS) versus foetal bovine serum (FBS) for growth of articular chondrocytes and formation of chondrocytefibrin constructs. Methods. Experiments with monolayer culture expansion of human articular chondroc...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BioMed Central Ltd.
2008
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/41837/ http://irep.iium.edu.my/41837/ http://irep.iium.edu.my/41837/1/AHS_versus_PHS_2008_MUNIRAH_ET_AL.pdf |
| Summary: | Purpose. To evaluate the effect of autologous human
serum (AHS) versus pooled human serum (PHS)
versus foetal bovine serum (FBS) for growth of
articular chondrocytes and formation of chondrocytefibrin
constructs.
Methods. Experiments with monolayer culture
expansion of human articular chondrocytes were
performed using basic culture media supplemented
with 10% AHS, PHS, or FBS. Growth kinetics
and specific phenotypic expression of the serially
expanded chondrocytes were evaluated. Largescale
culture expansion was used to obtain about 30
million cells to form chondrocyte-fibrin constructs.
All constructs were implanted subcutaneously at the
dorsum part of athymic nude mice for 8 weeks. The in
vivo constructs were evaluated using histological and
gene expression studies.
Results. The morphology of primary cultured
chondrocytes (P0) was polygonal and became more
elongated and larger after serial passages (P1, P2, and
P3). This was comparable for AHS, PHS, and FBS.
Total cell yields accumulated for AHS (28 million) and PHS (41 million) were significantly higher than those
for FBS (4 million). After 8 weeks of implantation,
in vivo chondrocyte-fibrin constructs demonstrated
a glistening white and firm texture, comparable to
normal hyaline cartilage. All constructs exhibited
histo-architectural characteristics of well-distributed
cartilage-isolated cells embedded within basophilic
ground substance. Presence of accumulated
proteoglycans cartilage-rich matrix was indicated
by positive orange-red Safranin O staining. During
monolayer culture expansion, collagen type II gene
expression was down-regulated, while collagen type
I gene expression was up-regulated. Collagen type
II—the specific chondrogenesis marker—was reexpressed
in the in vivo chondrocyte-fibrin construct.
Conclusion. AHS and PHS are better than FBS for in
vitro cultivation of human articular chondrocytes. |
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