Full
Outline
To compare different cryopreservation methods for lymphocytes and explore an optimized preservation protocol.
Cryoprotectant formulations [containing different concentrations of dimethyl sulfoxide (DMSO), fetal bovine serum (FBS), polyethylene glycol (PEG), and bovine serum albumin (BSA)] were screened for lymphocytes to identify two optimized formulations: experimental group 1 (C11): (7.5% DMSO + 10% FBS + 2.5% PEG + 2% BSA) and experimental group 2 (C21): (7.5% DMSO + 2.5% PEG + 2% BSA). On this basis, cryopreservation performance was compared between the two optimized cryoprotectant formulations paired with their respective optimal cooling rates (C11: −3 °C/min; C21: −1 °C/min) and the conventional cryopreservation method [control group (C0)]: (10% DMSO + 90% FBS) with gradient cooling. After 5 days of cryopreservation, cells were thawed to assess preservation quality indicators in terms of cell viability and functionality across groups.
The cell recovery rate at 0 h post-thawing in the control group (C0) (65.70 ± 1.06)% was lower than that in the experimental group 1 (C11) [(82.45 ± 3.27)%, t = −8.44, P < 0.05] and experimental group 2 (C21) [(83.96 ± 0.81)%, t = −23.72, P < 0.05]. The apoptosis rate at 0 h post-thawing in the control group (C0) (6.93 ± 0.55)% was higher than that in the experimental group 1 (C11) [(5.30% ± 0.56)%, t = 3.61, P < 0.05] and experimental group 2 (C21) [(4.93 ± 0.45)%, t = 4.87, P < 0.05]. Moreover, the differences between groups were statistically significant.
Based on multifactorial screening and optimization, the experimental group 2 using a serum-free cryoprotectant with a cooling rate of −1 °C/min outperforms the conventional cryopreservation method in terms of cell quality. This preservation protocol provides a scientific reference for the cryopreservation of lymphocytes.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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