IADR Abstract Archives
Adaptation of SHED Expanded in Foetal Bovine and Human Sera
Objectives: Paracrine factor profile within the patients’ body plays an important role in the regeneration process. Hence the adaptation of expanded allogenic MSCs in patients’ own serum prior to transplantation is an important phase. This study aimed to determine the viability and paracrine factor profile of stem cells from human extracted deciduous teeth (SHED) when adapted in individual human sera following expansion.
Methods: SHED (n=3) from passage 4 were expanded in foetal bovine serum (FBS-SHED) and pooled human serum (pHS-SHED) supplemented media up till passage 7. During the expansion, the proliferation of SHED was determined. Cells from these passages were then adapted in human serum (iHS) from 4 different male adults (21 to 35 years old) for a total of 120 hours. After 24hrs, cell viability were assessed using Prestoblue and paracrine factors profiled after 24 and 120hrs using Luminex based ProcartaPlex human cytokines 11 Plex immunoassay kit to elucidate the attainment of adaptation.
Results: Proliferation of SHED was significantly higher (p<.05), when expanded in pHS supplemented media compared FBS supplemented media. pHS-SHED maintained their higher proliferation rate compared to FBS-SHED, when they were adapted in iHS (n=4) supplemented media after 24hrs. During adaptation in iHS, FBS-SHED expressed significantly higher profile of SDF-1A (p<0.05) after 24hrs post-incubation compared to pHS-SHED. Similar observation was also found for HGF (p<0.01), LIF (p<0.05), PDGF-BB (p<0.05), SDF-1A (p<0.01) and IL-10 (p<0.05) when FBS-SHED was profiled after 120hrs post-incubation. These paracrine factors has been reported to be highly expressed by cells that showed signs of stress in oxygen, glucose and serum deprivation conditions.
Conclusions: The expression of paracrine factors by FBS-SHED during adaptation in iHS after 24 and 120hrs indicated that cells showed signs of stress when compare to pHS-SHED. This preliminary results indicated that 24hrs is sufficient for adaptation of pHS-SHED which could lead to better engraftment when cells at transplantation.
Methods: SHED (n=3) from passage 4 were expanded in foetal bovine serum (FBS-SHED) and pooled human serum (pHS-SHED) supplemented media up till passage 7. During the expansion, the proliferation of SHED was determined. Cells from these passages were then adapted in human serum (iHS) from 4 different male adults (21 to 35 years old) for a total of 120 hours. After 24hrs, cell viability were assessed using Prestoblue and paracrine factors profiled after 24 and 120hrs using Luminex based ProcartaPlex human cytokines 11 Plex immunoassay kit to elucidate the attainment of adaptation.
Results: Proliferation of SHED was significantly higher (p<.05), when expanded in pHS supplemented media compared FBS supplemented media. pHS-SHED maintained their higher proliferation rate compared to FBS-SHED, when they were adapted in iHS (n=4) supplemented media after 24hrs. During adaptation in iHS, FBS-SHED expressed significantly higher profile of SDF-1A (p<0.05) after 24hrs post-incubation compared to pHS-SHED. Similar observation was also found for HGF (p<0.01), LIF (p<0.05), PDGF-BB (p<0.05), SDF-1A (p<0.01) and IL-10 (p<0.05) when FBS-SHED was profiled after 120hrs post-incubation. These paracrine factors has been reported to be highly expressed by cells that showed signs of stress in oxygen, glucose and serum deprivation conditions.
Conclusions: The expression of paracrine factors by FBS-SHED during adaptation in iHS after 24 and 120hrs indicated that cells showed signs of stress when compare to pHS-SHED. This preliminary results indicated that 24hrs is sufficient for adaptation of pHS-SHED which could lead to better engraftment when cells at transplantation.