Benefit of trehalose preincubation prior to cryopreservation

Pawitan, Jeanne Adiwinata

doi:10.13181/mji.v25i3.1595

Cited by 1 publication

(1 citation statement)

References 15 publications

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“…A review on the failure of phase III clinical trial, which used over expanded cryopreserved mesenchymal stem cells to cure steroid resistant graft versus host disease, supposed that the failure was due to impaired immunomodulatory functions due to cryopreservation, and senescence due to overexpansion. 7,8 Therefore, for patients' safety and prospective comparisons between trial results, cells need to be characterized, and their function and senescent profile need to be analyzed.…”

Section: Introductionmentioning

confidence: 99%

Effect of Cryopreservation and cumulative population doublings on Senescence of Umbilical Cord Mesenchymal Stem Cells

Pawitan¹,

Goei²,

Liem³

et al. 2017

IJPTR

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: Background: Multiple harvest explant method (MHEM) derived umbilical cord mesenchymal stem cells (UC-MSCs) was shown to undergo senescent beginning at passage-10 (P-10). However, for the same cells, there are no senescent data after cryopreservation and passage. Therefore, this study aimed to analyze the senescent profile of cryopreserved MHEM derived UC-MSCs after serial passages. Methods: MHEM derived UC-MSCs were isolated and cultured in platelet lysate (PL) containing medium as described previously. The cells were cryopreserved at passage-1 (P-1) in 10% dimethyl sulfoxide (DMSO) and 10% PL containing alpha minimal essential medium (αMEM). Cell density at cryopreservation was 500 000 cells/mL. After one month, the cells were thawed and recultured until P-8 in six 12-well plate. At 80-90% confluent, two wells were harvested and the cells were recultured into six wells, and the four remaining wells were subjected to senescent (β-galactosidase) staining, which was done for all passages. Random photographs were taken from all stained wells, and senescent percentage was recorded. Results: No senescent cells were observed at P-2. Senescent cells began to appear at P-3. Percentage (mean ± SD) of senescent cells from P-3 through P-8 were 0.04 ± 0.02, 1.18 ± 1.82, 0.14 ± 0.18, 0.49 ± 0.00, 0.58 ±0.91, and 0.07 ± 0.07, respectively. Results: No senescent cells were observed at P-2. Senescent cells began to appear at P-3. Percentage (mean ± SD) of senescent cells from P-3 through P-8 were 0.04 ± 0.02, 1.18 ± 1.82, 0.14 ± 0.18, 0.49 ± 0.00, 0.58 ±0.91, and 0.07 ± 0.07, respectively. Conclusion: No senescent cells were observed at P-2 (cumulative population doubling [CPD] > 9.38). Senescent cells began to appear at P-3 (CPD > 14.06), but in all passages until P-8 (CPD >34.34) the senescent percentage was below 5%.

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Section: Introductionmentioning

confidence: 99%