Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes

Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes

Traditionally, foetal bovine serum (FBS) is used as a serum supplement for stem cell expansion in vitro. However, it is associated with xenoimmunisation and the transmission of animal pathogens, which may cause harm to stem cell recipients. As a safer alternative, human platelet lysate (HPL) has bee...

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Main Authors: Prana Hardinata Budi Harto, Muhammad Hanif Mahmud, Aisya Hanim Othman, Nurulain Hanani Ngadenin, Nur Saihah Mohd Azahar, Muhammad Najib Fathi Hassan, Nor Hamdan Mohd Yahaya, Rizal Abdul Rani, Leong, Chooi Fun, Ng, Min Hwei, Law, Jia Xian
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2019
Online Access:http://journalarticle.ukm.my/14389/
http://journalarticle.ukm.my/14389/
http://journalarticle.ukm.my/14389/1/12%20Prana%20Hardinata.pdf
id ukm-14389
recordtype eprints
spelling ukm-143892020-03-10T02:32:23Z http://journalarticle.ukm.my/14389/ Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes Prana Hardinata Budi Harto, Muhammad Hanif Mahmud, Aisya Hanim Othman, Nurulain Hanani Ngadenin, Nur Saihah Mohd Azahar, Muhammad Najib Fathi Hassan, Nor Hamdan Mohd Yahaya, Rizal Abdul Rani, Leong, Chooi Fun Ng, Min Hwei Law, Jia Xian Traditionally, foetal bovine serum (FBS) is used as a serum supplement for stem cell expansion in vitro. However, it is associated with xenoimmunisation and the transmission of animal pathogens, which may cause harm to stem cell recipients. As a safer alternative, human platelet lysate (HPL) has been introduced for propagating stem cells. Chondrocytes are expanded in vitro for cartilage repair via autologous chondrocyte implantation (ACI). In this study, we compare the efficacy of HPL prepared from expired platelet concentrates with that of FBS for promoting the proliferation and maintenance of the chondrogenic markers of primary human chondrocytes expanded in vitro. Chondrocytes were cultured in F12:Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% HPL, 10% HPL and 10% FBS. The cell morphology, viability and growth rate were examined from passage 1 (P1) to P3. RNA was isolated from P3 cells for quantitative polymerase chain reaction (qPCR) to determine the gene expression level of the chondrogenic, dedifferentiation and hypertrophic markers. HPL promoted chondrocyte proliferation without compromising cell viability. In addition, the chondrocytes cultured with HPL were smaller. However, HPL failed to maintain the chondrogenic markers, except SOX 9 (SRY-box transcription factor 9), which was upregulated, but not significantly. Nonetheless, HPL also suppressed the expression of type X collagen (Col X), a chondrocyte hypertrophic marker. In summary, we demonstrate the benefits of HPL supplementation in human chondrocyte culture, where it enhances cell proliferation and suppresses chondrocyte hypertrophy. In the future, HPL can be used for the large-scale expansion of chondrocytes for ACI. Penerbit Universiti Kebangsaan Malaysia 2019-10 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/14389/1/12%20Prana%20Hardinata.pdf Prana Hardinata Budi Harto, and Muhammad Hanif Mahmud, and Aisya Hanim Othman, and Nurulain Hanani Ngadenin, and Nur Saihah Mohd Azahar, and Muhammad Najib Fathi Hassan, and Nor Hamdan Mohd Yahaya, and Rizal Abdul Rani, and Leong, Chooi Fun and Ng, Min Hwei and Law, Jia Xian (2019) Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes. Sains Malaysiana, 48 (10). pp. 2169-2176. ISSN 0126-6039 http://www.ukm.my/jsm/malay_journals/jilid48bil10_2019/KandunganJilid48Bil10_2019.htm
repository_type Digital Repository
institution_category Local University
institution Universiti Kebangasaan Malaysia
building UKM Institutional Repository
collection Online Access
language English
description Traditionally, foetal bovine serum (FBS) is used as a serum supplement for stem cell expansion in vitro. However, it is associated with xenoimmunisation and the transmission of animal pathogens, which may cause harm to stem cell recipients. As a safer alternative, human platelet lysate (HPL) has been introduced for propagating stem cells. Chondrocytes are expanded in vitro for cartilage repair via autologous chondrocyte implantation (ACI). In this study, we compare the efficacy of HPL prepared from expired platelet concentrates with that of FBS for promoting the proliferation and maintenance of the chondrogenic markers of primary human chondrocytes expanded in vitro. Chondrocytes were cultured in F12:Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% HPL, 10% HPL and 10% FBS. The cell morphology, viability and growth rate were examined from passage 1 (P1) to P3. RNA was isolated from P3 cells for quantitative polymerase chain reaction (qPCR) to determine the gene expression level of the chondrogenic, dedifferentiation and hypertrophic markers. HPL promoted chondrocyte proliferation without compromising cell viability. In addition, the chondrocytes cultured with HPL were smaller. However, HPL failed to maintain the chondrogenic markers, except SOX 9 (SRY-box transcription factor 9), which was upregulated, but not significantly. Nonetheless, HPL also suppressed the expression of type X collagen (Col X), a chondrocyte hypertrophic marker. In summary, we demonstrate the benefits of HPL supplementation in human chondrocyte culture, where it enhances cell proliferation and suppresses chondrocyte hypertrophy. In the future, HPL can be used for the large-scale expansion of chondrocytes for ACI.
format Article
author Prana Hardinata Budi Harto,
Muhammad Hanif Mahmud,
Aisya Hanim Othman,
Nurulain Hanani Ngadenin,
Nur Saihah Mohd Azahar,
Muhammad Najib Fathi Hassan,
Nor Hamdan Mohd Yahaya,
Rizal Abdul Rani,
Leong, Chooi Fun
Ng, Min Hwei
Law, Jia Xian
spellingShingle Prana Hardinata Budi Harto,
Muhammad Hanif Mahmud,
Aisya Hanim Othman,
Nurulain Hanani Ngadenin,
Nur Saihah Mohd Azahar,
Muhammad Najib Fathi Hassan,
Nor Hamdan Mohd Yahaya,
Rizal Abdul Rani,
Leong, Chooi Fun
Ng, Min Hwei
Law, Jia Xian
Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
author_facet Prana Hardinata Budi Harto,
Muhammad Hanif Mahmud,
Aisya Hanim Othman,
Nurulain Hanani Ngadenin,
Nur Saihah Mohd Azahar,
Muhammad Najib Fathi Hassan,
Nor Hamdan Mohd Yahaya,
Rizal Abdul Rani,
Leong, Chooi Fun
Ng, Min Hwei
Law, Jia Xian
author_sort Prana Hardinata Budi Harto,
title Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
title_short Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
title_full Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
title_fullStr Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
title_full_unstemmed Human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
title_sort human platelet lysate promotes proliferation but fails to maintain chondrogenic markers of chondrocytes
publisher Penerbit Universiti Kebangsaan Malaysia
publishDate 2019
url http://journalarticle.ukm.my/14389/
http://journalarticle.ukm.my/14389/
http://journalarticle.ukm.my/14389/1/12%20Prana%20Hardinata.pdf
first_indexed 2023-09-18T20:06:58Z
last_indexed 2023-09-18T20:06:58Z
_version_ 1777407222839508992

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