Autologous versus pooled human serum for articular chondrocyte growth.

S. Munirah, Ruszymah Idrus, O. C. Samsudin, Md. Yusoff Badrul Akmal Hisham, Azmi Baharudin, B. S. Aminuddin

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

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. Large-scale 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 re-expressed in the in vivo chondrocyte-fibrin construct. CONCLUSION: AHS and PHS are better than FBS for in vitro cultivation of human articular chondrocytes.

Original languageEnglish
Pages (from-to)220-229
Number of pages10
JournalJournal of orthopaedic surgery (Hong Kong)
Volume16
Issue number2
Publication statusPublished - Aug 2008

Fingerprint

Chondrocytes
Joints
Growth
Serum
Fibrin
Collagen Type II
Gene Expression
Nude Mice
Cartilage
Hyaline Cartilage
Serial Passage
Chondrogenesis
Proteoglycans
Collagen Type I
Culture Media

ASJC Scopus subject areas

  • Surgery

Cite this

Autologous versus pooled human serum for articular chondrocyte growth. / Munirah, S.; Idrus, Ruszymah; Samsudin, O. C.; Badrul Akmal Hisham, Md. Yusoff; Baharudin, Azmi; Aminuddin, B. S.

In: Journal of orthopaedic surgery (Hong Kong), Vol. 16, No. 2, 08.2008, p. 220-229.

Research output: Contribution to journalArticle

@article{d45298792f6f44af8f59d273d97760a8,
title = "Autologous versus pooled human serum for articular chondrocyte growth.",
abstract = "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. Large-scale 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 re-expressed in the in vivo chondrocyte-fibrin construct. CONCLUSION: AHS and PHS are better than FBS for in vitro cultivation of human articular chondrocytes.",
author = "S. Munirah and Ruszymah Idrus and Samsudin, {O. C.} and {Badrul Akmal Hisham}, {Md. Yusoff} and Azmi Baharudin and Aminuddin, {B. S.}",
year = "2008",
month = "8",
language = "English",
volume = "16",
pages = "220--229",
journal = "Journal of orthopaedic surgery (Hong Kong)",
issn = "1022-5536",
publisher = "Hong Kong Academy of Medicine Press",
number = "2",

}

TY - JOUR

T1 - Autologous versus pooled human serum for articular chondrocyte growth.

AU - Munirah, S.

AU - Idrus, Ruszymah

AU - Samsudin, O. C.

AU - Badrul Akmal Hisham, Md. Yusoff

AU - Baharudin, Azmi

AU - Aminuddin, B. S.

PY - 2008/8

Y1 - 2008/8

N2 - 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. Large-scale 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 re-expressed in the in vivo chondrocyte-fibrin construct. CONCLUSION: AHS and PHS are better than FBS for in vitro cultivation of human articular chondrocytes.

AB - 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. Large-scale 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 re-expressed in the in vivo chondrocyte-fibrin construct. CONCLUSION: AHS and PHS are better than FBS for in vitro cultivation of human articular chondrocytes.

UR - http://www.scopus.com/inward/record.url?scp=59649129503&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=59649129503&partnerID=8YFLogxK

M3 - Article

C2 - 18725677

AN - SCOPUS:59649129503

VL - 16

SP - 220

EP - 229

JO - Journal of orthopaedic surgery (Hong Kong)

JF - Journal of orthopaedic surgery (Hong Kong)

SN - 1022-5536

IS - 2

ER -