Pukana Jayaraman1,2, Vijayendran Govindasamy3, Nareshwaran Gnanasegaran1,4, Wijenthiran Kunasekaran1,2, Punitha Vasanthan1,2, Sabri Musa5, Noor Hayaty Abu Kasim4. 1. GMP compliant Stem Cells Laboratory, Hygieia Innovation Sdn. Bhd, Federal Territory of Putrajaya, Lot 1G-2G, Lanai Complex, No.2, Persiaran Seri Perdana, Precinct 10, 62250, Putrajaya, Malaysia. 2. Department of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, University of Malaya, 50603, Kuala Lumpur, Malaysia. 3. GMP compliant Stem Cells Laboratory, Hygieia Innovation Sdn. Bhd, Federal Territory of Putrajaya, Lot 1G-2G, Lanai Complex, No.2, Persiaran Seri Perdana, Precinct 10, 62250, Putrajaya, Malaysia. vijay@hygieiainnovation.com. 4. Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, 50603, Kuala Lumpur, Malaysia. 5. Department of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, University of Malaya, 50603, Kuala Lumpur, Malaysia. sabrim@um.edu.my.
Abstract
BACKGROUND AND OBJECTIVES: Long-term culture system is used to prevent the impediment of insufficient cells and is good for low starting materials such as dental pulp or periodontal ligament. In general, although cell viability and functionality are the most common aspects taken into consideration in culturing cells for a long term, they may not truly represent the biological state of the cells. Hence, we explored the behaviour of another important aspect which is the immune properties in long-term cultured cells. METHODS: Dental pulp stem cells from deciduous (SHED; n = 3) and permanent (DPSCs; n = 3) teeth as well as periodontal ligament stem cells (PDLSCs; n = 3) were cultured under identical culture condition. The immune properties of each cell lines were profiled at passage 2 [P2] and passage 9 [P9] as early and late passages, respectively. This was further validated at the protein level using the Luminex platform. RESULTS: A major shift of genes was noticed at P9 with SHED being the highest. SHED cultured at P9 displayed many genes representing pathogen recognition (P < 0.001), immune signalling (P < 0.001, pro-inflammatory (P < 0.001), anti-inflammatory (P < 0.001) and immune-related growth and stimulation factor (P < 0.001) as compared to DPSCs and PDLSCs. Surprisingly, SHED also expressed many cytotoxicity genes (P < 0.001). CONCLUSIONS: Communally, instabilities of immune genes from our findings suggest that long-term cultured cells may not be feasible for transplantation purposes. CLINICAL RELEVANCE: A complete biological characterization covering all major aspects including immune properties should be made as prerequisite criteria prior to the use of long-term cultured stem cells in clinical settings.
BACKGROUND AND OBJECTIVES: Long-term culture system is used to prevent the impediment of insufficient cells and is good for low starting materials such as dental pulp or periodontal ligament. In general, although cell viability and functionality are the most common aspects taken into consideration in culturing cells for a long term, they may not truly represent the biological state of the cells. Hence, we explored the behaviour of another important aspect which is the immune properties in long-term cultured cells. METHODS: Dental pulp stem cells from deciduous (SHED; n = 3) and permanent (DPSCs; n = 3) teeth as well as periodontal ligament stem cells (PDLSCs; n = 3) were cultured under identical culture condition. The immune properties of each cell lines were profiled at passage 2 [P2] and passage 9 [P9] as early and late passages, respectively. This was further validated at the protein level using the Luminex platform. RESULTS: A major shift of genes was noticed at P9 with SHED being the highest. SHED cultured at P9 displayed many genes representing pathogen recognition (P < 0.001), immune signalling (P < 0.001, pro-inflammatory (P < 0.001), anti-inflammatory (P < 0.001) and immune-related growth and stimulation factor (P < 0.001) as compared to DPSCs and PDLSCs. Surprisingly, SHED also expressed many cytotoxicity genes (P < 0.001). CONCLUSIONS: Communally, instabilities of immune genes from our findings suggest that long-term cultured cells may not be feasible for transplantation purposes. CLINICAL RELEVANCE: A complete biological characterization covering all major aspects including immune properties should be made as prerequisite criteria prior to the use of long-term cultured stem cells in clinical settings.
Authors: R Vecchiatini; L Penolazzi; E Lambertini; M Angelozzi; C Morganti; S Mazzitelli; L Trombelli; C Nastruzzi; R Piva Journal: J Periodontal Res Date: 2014-09-23 Impact factor: 4.419
Authors: Trivia P Frazier; Jeffrey M Gimble; Jessica W Devay; Hugh A Tucker; Ernest S Chiu; Brian G Rowan Journal: BMC Cell Biol Date: 2013-08-07 Impact factor: 4.241
Authors: Sueli Patricia Harumi Miyagi de Cara; Clarice Silvia Taemi Origassa; Fernando de Sá Silva; Maria Stella N A Moreira; Danilo Candido de Almeida; Ana Clara Fagundes Pedroni; Giovanna Lopes Carvalho; Diego Pulzatto Cury; Niels Olsen Saraiva Câmara; Márcia Martins Marques Journal: Heliyon Date: 2019-04-28
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