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Autologous serum supplement favours in vitro regenerative paracrine factors synthesis

Objectives: Foetal bovine serum (FBS) is often the serum supplement of choice for in vitro human cell culture. This study compares the effect of FBS and autologous human serum (AuHS) supplement in human peripheral blood mononuclear cell (PBMC) culture to prepare secretome. Materials and methods: The...

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Main Authors: Haque, N., Abu Kasim, N.H., Abu Kassim, N.L., Rahman, M.T.
Format: Article
Language:English
Published: Blackwell Publishing 2017
Subjects:
Practice of dentistry. Dental economics
Online Access:http://eprints.um.edu.my/17751/1/Autologous_serum_supplement_favours_in_vitro_regenerative_paracrine.pdf
http://eprints.um.edu.my/17751/
http://onlinelibrary.wiley.com/doi/10.1111/cpr.12354/abstract;jsessionid=73A17491F64437D81D45D959E8319A95.f04t02
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spelling my.um.eprints.177512017-09-13T03:58:45Z http://eprints.um.edu.my/17751/ Autologous serum supplement favours in vitro regenerative paracrine factors synthesis Haque, N. Abu Kasim, N.H. Abu Kassim, N.L. Rahman, M.T. Practice of dentistry. Dental economics Objectives: Foetal bovine serum (FBS) is often the serum supplement of choice for in vitro human cell culture. This study compares the effect of FBS and autologous human serum (AuHS) supplement in human peripheral blood mononuclear cell (PBMC) culture to prepare secretome. Materials and methods: The PBMC (n = 7) were cultured either in RPMI-1640 containing L-glutamine and 50 units/ml Penicillin-Streptomycin (BM) or in BM with either AuHS or FBS. Viability, proliferation and differentiation of PBMC were evaluated. Paracrine factors present in the secretomes (n = 6) were analysed using ProcartaPlex Human Cytokine panel (17 plex). Ingenuity Pathway Analysis (IPA) was performed to predict activation or inhibition of biological functions related to tissue regeneration. Results: The viability of PBMC that were cultured with FBS supplement was significantly reduced at 96 h compared to those at 0 and 24 h (P < .05). While the reduction of the viability of PBMC that were cultured with AuHS supplement was not significantly different compared to those at 0 and 24 h. The FBS secretomes prepared at 24 h was found to contain significantly higher amount of EGF (P < .05) compared to that in AuHS or BM secretome. The AuHS secretomes contained significantly higher amount of HGF at 24 (P < .05) and 96 h (P < .01), and VEGF-A at 24 h (P < .05) compared to those in the FBS secretomes. SDF-1 was not detected in the FBS secretomes prepared at either 24 or 96 hours. Double immunocytochemical staining revealed a marked increase in co-localization of SDF-1 and its receptor in PBMC that were cultured with AuHS supplement compared to that cultured with FBS supplement. Conclusion: In secretome preparation, AuHS supplement favours synthesis of paracrine factors that are needed for regenerative therapy. Blackwell Publishing 2017 Article PeerReviewed application/pdf en http://eprints.um.edu.my/17751/1/Autologous_serum_supplement_favours_in_vitro_regenerative_paracrine.pdf Haque, N. and Abu Kasim, N.H. and Abu Kassim, N.L. and Rahman, M.T. (2017) Autologous serum supplement favours in vitro regenerative paracrine factors synthesis. Cell Proliferation, 50 (4). e12354. ISSN 0960-7722 http://onlinelibrary.wiley.com/doi/10.1111/cpr.12354/abstract;jsessionid=73A17491F64437D81D45D959E8319A95.f04t02 doi:10.1111/cpr.12354
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
language English
topic Practice of dentistry. Dental economics
spellingShingle Practice of dentistry. Dental economics
Haque, N.
Abu Kasim, N.H.
Abu Kassim, N.L.
Rahman, M.T.
Autologous serum supplement favours in vitro regenerative paracrine factors synthesis
description Objectives: Foetal bovine serum (FBS) is often the serum supplement of choice for in vitro human cell culture. This study compares the effect of FBS and autologous human serum (AuHS) supplement in human peripheral blood mononuclear cell (PBMC) culture to prepare secretome. Materials and methods: The PBMC (n = 7) were cultured either in RPMI-1640 containing L-glutamine and 50 units/ml Penicillin-Streptomycin (BM) or in BM with either AuHS or FBS. Viability, proliferation and differentiation of PBMC were evaluated. Paracrine factors present in the secretomes (n = 6) were analysed using ProcartaPlex Human Cytokine panel (17 plex). Ingenuity Pathway Analysis (IPA) was performed to predict activation or inhibition of biological functions related to tissue regeneration. Results: The viability of PBMC that were cultured with FBS supplement was significantly reduced at 96 h compared to those at 0 and 24 h (P < .05). While the reduction of the viability of PBMC that were cultured with AuHS supplement was not significantly different compared to those at 0 and 24 h. The FBS secretomes prepared at 24 h was found to contain significantly higher amount of EGF (P < .05) compared to that in AuHS or BM secretome. The AuHS secretomes contained significantly higher amount of HGF at 24 (P < .05) and 96 h (P < .01), and VEGF-A at 24 h (P < .05) compared to those in the FBS secretomes. SDF-1 was not detected in the FBS secretomes prepared at either 24 or 96 hours. Double immunocytochemical staining revealed a marked increase in co-localization of SDF-1 and its receptor in PBMC that were cultured with AuHS supplement compared to that cultured with FBS supplement. Conclusion: In secretome preparation, AuHS supplement favours synthesis of paracrine factors that are needed for regenerative therapy.
format Article
author Haque, N.
Abu Kasim, N.H.
Abu Kassim, N.L.
Rahman, M.T.
author_facet Haque, N.
Abu Kasim, N.H.
Abu Kassim, N.L.
Rahman, M.T.
author_sort Haque, N.
title Autologous serum supplement favours in vitro regenerative paracrine factors synthesis
title_short Autologous serum supplement favours in vitro regenerative paracrine factors synthesis
title_full Autologous serum supplement favours in vitro regenerative paracrine factors synthesis
title_fullStr Autologous serum supplement favours in vitro regenerative paracrine factors synthesis
title_full_unstemmed Autologous serum supplement favours in vitro regenerative paracrine factors synthesis
title_sort autologous serum supplement favours in vitro regenerative paracrine factors synthesis
publisher Blackwell Publishing
publishDate 2017
url http://eprints.um.edu.my/17751/1/Autologous_serum_supplement_favours_in_vitro_regenerative_paracrine.pdf
http://eprints.um.edu.my/17751/
http://onlinelibrary.wiley.com/doi/10.1111/cpr.12354/abstract;jsessionid=73A17491F64437D81D45D959E8319A95.f04t02
_version_ 1643690508659195904
score 13.211869

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