1 research outputs found
Analysis of the structural and functional state of human cord blood hematopoietic progenitor cells after cryopreservation with DMSO and antioxidants and transfer to conditions close to physiological
Π£ ΡΠΎΠ±ΠΎΡΡ ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΎ ΠΏΠΎΡΡΠ²Π½ΡΠ»ΡΠ½ΠΈΠΉ Π°Π½Π°Π»ΡΠ· Π²ΠΏΠ»ΠΈΠ²Ρ ΠΊΡΡΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ½ΠΈΡ
ΡΡΠΌΡΡΠ΅ΠΉ, ΡΠΎ ΠΌΡΡΡΡΡΡ ΡΡΠ·Π½Ρ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΡΡ ΠΏΡΠΎΠ½ΠΈΠΊΠ°ΡΡΠΎΠ³ΠΎ ΠΊΡΡΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ° ΠΠΠ‘Π ΡΠ° Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΡΠ², ΠΏΡΠΈ ΠΊΡΡΠΎΠΊΠΎΠ½ΡΠ΅ΡΠ²ΡΠ²Π°Π½Π½Ρ Π³Π΅ΠΌΠΎΠΏΠΎΠ΅ΡΠΈΡΠ½ΠΈΡ
ΠΏΡΠΎΠ³Π΅Π½ΡΡΠΎΡΠ½ΠΈΡ
ΠΊΠ»ΡΡΠΈΠ½ ΠΊΠΎΡΠ΄ΠΎΠ²ΠΎΡ ΠΊΡΠΎΠ²Ρ Π»ΡΠ΄ΠΈΠ½ΠΈ ΡΠ° ΠΏΡΡΠ»Ρ ΠΏΠ΅ΡΠ΅Π½Π΅ΡΠ΅Π½Π½Ρ ΡΡ
Π΄ΠΎ ΡΠΌΠΎΠ², Π½Π°Π±Π»ΠΈΠΆΠ΅Π½ΠΈΡ
Π΄ΠΎ ΡΡΠ·ΡΠΎΠ»ΠΎΠ³ΡΡΠ½ΠΈΡ
. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΠΎ Π΄ΠΎΠ΄Π°Π²Π°Π½Π½Ρ Π΄ΠΎ ΠΊΡΡΠΎΠ·Π°Ρ
ΠΈΡΠ½ΠΎΠ³ΠΎ ΡΠ΅ΡΠ΅Π΄ΠΎΠ²ΠΈΡΠ° Π°ΡΠΊΠΎΡΠ±ΡΠ½ΠΎΠ²ΠΎΡ ΠΊΠΈΡΠ»ΠΎΡΠΈ, N-Π°ΡΠ΅ΡΠΈΠ»-L-ΡΠΈΡΡΠ΅ΡΠ½Π° Π°Π±ΠΎ Π³Π»ΡΡΠ°ΡΡΠΎΠ½Π° ΡΠΏΡΠΈΡΡ ΠΏΡΠ΄Π²ΠΈΡΠ΅Π½Π½Ρ ΠΏΠΎΠΊΠ°Π·Π½ΠΈΠΊΡΠ² Π·Π±Π΅ΡΠ΅ΠΆΠ΅Π½ΠΎΡΡΡ ΡΠ° ΠΆΠΈΡΡΡΠ·Π΄Π°ΡΠ½ΠΎΡΡΡ Π³Π΅ΠΌΠΎΠΏΠΎΠ΅ΡΠΈΡΠ½ΠΈΡ
ΠΏΡΠΎΠ³Π΅Π½ΡΡΠΎΡΠ½ΠΈΡ
ΠΊΠ»ΡΡΠΈΠ½ ΠΏΡΡΠ»Ρ ΠΏΠ΅ΡΠ΅Π½Π΅ΡΠ΅Π½Π½Ρ ΡΡ
Π΄ΠΎ ΡΠΌΠΎΠ², ΡΠΎ ΠΌΠΎΠ΄Π΅Π»ΡΡΡΡ ΡΡΠ·ΡΠΎΠ»ΠΎΠ³ΡΡΠ½Ρ. ΠΠ°ΠΉΠ±ΡΠ»ΡΡΠΈΠΉ ΡΠΈΡΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ½ΠΈΠΉ Π΅ΡΠ΅ΠΊΡ ΡΠΏΠΎΡΡΠ΅ΡΡΠ³Π°ΡΡΡΡΡ Π² ΠΏΡΠΎΠ±Π°Ρ
, ΠΊΡΡΠΎΠΊΠΎΠ½ΡΠ΅ΡΠ²ΠΎΠ²Π°Π½ΠΈΡ
ΡΠ· 7,5 Ρ 10% ΠΠΠ‘Π Π· Π΄ΠΎΠ΄Π°Π²Π°Π½Π½ΡΠΌ Π³Π»ΡΡΠ°ΡΡΠΎΠ½Ρ Π² ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΡΡ 1 ΡΠ° 3 ΠΌΠ, Π΄Π΅ Π·Π±Π΅ΡΡΠ³Π°ΡΡΡΡΡ Π΄ΠΎ 80% Π³Π΅ΠΌΠΎΠΏΠΎΠ΅ΡΠΈΡΠ½ΠΈΡ
ΠΊΠ»ΡΡΠΈΠ½ Π² ΠΆΠΈΡΡΡΠ·Π΄Π°ΡΠ½ΠΎΠΌΡ ΡΡΠ°Π½Ρ, ΠΏΠΎΡΡΠ²Π½ΡΠ½ΠΎ Π· Π΄Π²ΠΎΠΌΠ° ΡΠ½ΡΠΈΠΌΠΈ Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠ°ΠΌΠΈ Π² ΠΎΠΏΡΠΈΠΌΠ°Π»ΡΠ½ΠΈΡ
ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΡΡΡ
, ΡΠΊΡ Π·Π°Π±Π΅Π·ΠΏΠ΅ΡΡΡΡΡ ΠΆΠΈΡΡΡΠ·Π΄Π°ΡΠ½ΡΡΡΡ ΠΊΠ»ΡΡΠΈΠ½ Π΄ΠΎ 70%; Π ΡΠ°Π±ΠΎΡΠ΅ ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ ΡΡΠ°Π²Π½ΠΈΡΠ΅Π»ΡΠ½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· Π²Π»ΠΈΡΠ½ΠΈΡ ΠΊΡΠΈΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ½ΡΡ
ΡΠΌΠ΅ΡΠ΅ΠΉ, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠΈΡ
ΡΠ°Π·Π»ΠΈΡΠ½ΡΠ΅ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ Π½Π΅ΠΏΡΠΎΠ½ΠΈΠΊΠ°ΡΡΠ΅Π³ΠΎ ΠΊΡΠΈΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ° ΠΠΠ‘Π ΠΈ Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠΎΠ², ΠΏΡΠΈ ΠΊΡΠΈΠΎΠΊΠΎΠ½ΡΠ΅ΡΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΠΈ Π³Π΅ΠΌΠΎΠΏΠΎΡΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΏΡΠΎΠ³Π΅Π½ΠΈΡΠΎΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΠΊΠΎΡΠ΄ΠΎΠ²ΠΎΠΉ ΠΊΡΠΎΠ²ΠΈ ΠΈ ΠΏΠΎΡΠ»Π΅ ΠΏΠ΅ΡΠ΅Π½ΠΎΡΠ° ΠΈΡ
Π² ΡΡΠ»ΠΎΠ²ΠΈΡ, ΠΏΡΠΈΠ±Π»ΠΈΠΆΠ΅Π½Π½ΡΠ΅ ΠΊ ΡΠΈΠ·ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΠΌ. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ Π΄ΠΎΠ±Π°Π²Π»Π΅Π½ΠΈΠ΅ ΠΊ ΠΊΡΠΈΠΎΠ·Π°ΡΠΈΡΠ½ΠΎΠΉ ΡΡΠ΅Π΄Π΅ Π°ΡΠΊΠΎΡΠ±ΠΈΠ½ΠΎΠ²ΠΎΠΉ ΠΊΠΈΡΠ»ΠΎΡΡ, N-Π°ΡΠ΅ΡΠΈΠ»-L-ΡΠΈΡΡΠ΅ΠΈΠ½Π° ΠΈΠ»ΠΈ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° ΡΠΏΠΎΡΠΎΠ±ΡΡΠ²ΡΠ΅Ρ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ΠΉ ΡΠΎΡ
ΡΠ°Π½Π½ΠΎΡΡΠΈ ΠΈ ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΠΈ Π³Π΅ΠΌΠΎΠΏΠΎΡΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΏΡΠΎΠ³Π΅Π½ΠΈΡΠΎΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΠΎΡΠ»Π΅ ΠΏΠ΅ΡΠ΅Π½ΠΎΡΠ° ΠΈΡ
Π² ΡΡΠ»ΠΎΠ²ΠΈΡ, ΠΌΠΎΠ΄Π΅Π»ΠΈΡΡΡΡΠΈΠ΅ ΡΠΈΠ·ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΠ΅. ΠΠ°ΠΈΠ±ΠΎΠ»ΡΡΠΈΠΉ ΡΠΈΡΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ½ΡΠΉ ΡΡΡΠ΅ΠΊΡ Π½Π°Π±Π»ΡΠ΄Π°Π΅ΡΡΡ Π² ΠΏΡΠΎΠ±Π°Ρ
, ΠΊΡΠΈΠΎΠΊΠΎΠ½ΡΠ΅ΡΠ²ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Ρ 7,5 ΠΈ 10% ΠΠΠ‘Π Ρ Π΄ΠΎΠ±Π°Π²Π»Π΅Π½ΠΈΠ΅ΠΌ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° Π² ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ 1 ΠΈ 3 ΠΌΠ, Π³Π΄Π΅ ΡΠΎΡ
ΡΠ°Π½ΡΠ΅ΡΡΡ Π΄ΠΎ 80% Π³Π΅ΠΌΠΎΠΏΠΎΡΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π² ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΠΌ ΡΠΎΡΡΠΎΡΠ½ΠΈΠΈ, ΠΏΠΎ ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ Ρ Π΄Π²ΡΠΌΡ Π΄ΡΡΠ³ΠΈΠΌΠΈ Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠ°ΠΌΠΈ Π² ΠΎΠΏΡΠΈΠΌΠ°Π»ΡΠ½ΡΡ
ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΡΡ
, ΠΊΠΎΡΠΎΡΡΠ΅ ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°ΡΡ ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΡ ΡΠΎΠ»ΡΠΊΠΎ Π΄ΠΎ 70% ΠΊΠ»Π΅ΡΠΎΠΊ; The use of cord blood (CB) hematopoietic progenitor cells (HPCs) for the past 15 years has been firmly establishing in practical medicine as an effective treatment for diseases of various origins. In this regard, it remains relevant for the development of protocols for cells storage at low temperature. Most of them are based on the use of penetrating cryoprotectant DMSO in effective concentrations. However, they do not take into account that during cryopreservation HPCs are subjected to a significant stress, resulting in the decrease of the antioxidant system work, disruption of the membrane structure and the glutathione release from the organelles, which leads to an increase in the level of ROS in cells after transferring them to the bloodstream and the development of lipid peroxidation. In addition, there are suggestions that in the initial period after warming, the level of lipid peroxidation is limited by the structural antioxidants of the glutathione peroxidase system. In this regard, to increase the efficiency of cryopreservation of cord blood preparations, an important task is not only to evaluate them immediately after thawing, but also to determine the delayed survival of cells after transport them to conditions close to physiological. The addition of antioxidants to the cryoprotective medium can improve the preservation rate and viability of HPCs both immediately after cryopreservation and after transfer to conditions that are close to physiological. Based on this, the aim of the work was to analysis of the structural and functional state of human cord blood hematopoietic progenitor cells after cryopreservation with cryoprotectant DMSO and antioxidants and transfer to conditions close to physiological The efficacy of ascorbic acid, N-acetyl-L-cysteine and glutathione application in combination with DMSO in cryopreservation of HPCs has been evaluated. It has been shown that in the process of cryopreservation with DMSO and after transferring to conditions that are close to physiological, the preservation and viability of HPCs decreases on 20% in the samples, cryopreserved with 7.5-10% DMSO. One of the reasons for this may be the accumulation of reactive oxygen species (ROS) in cells during freezing. The addition of antioxidants to the cryoprotective medium can significantly increase the stability of the HPCs against the effects of cryopreservation factors and improve the preservation and viability indices. A comparative analysis of antioxidants revealed that ascorbic acid at concentrations of 0.1 and 0.15 mM and N-acetyl-L-cysteine (10 and 15 mM) in combination with 7.5% and 10% DMSO increased the number of preserved viable HPCs up to 70% in comparison to the samples without adding the antioxidants. Addition of glutathione at a concentration of 1 and 3 mM to cryoprotective medium with 7.5% and 10% DMSO was able to maintain a viable state of up to 80% of the HPCs. This may be due to the fact that glutathione reduced the number of cells with excess content of ROS after transferring the cells to conditions that are close to physiological