Abstract
The performed comparative analysis of structural and functional status of fetal liver (FL) hematopoietic stem cells (HSCs) of 14 gestation days and adult bone marrow (BM) of 4-month-old CBA/H mice demonstrated not only the differences of their initial qualitative and quantitative characteristics but also different responses to the effect of cryopreservation factors. Either FL or BM cells were frozen according to various regimens under protection of 5 and 10% dimethyl sulfoxide. To estimate structural and functional characteristics of HSCs derived from different sources there were used cytofluorimetry (content of CD34+СD38– cells), methods of colony formation in vivo (CFUs) and in vitro (CFU-GM). It has been established that certain cryopreservation regimens are able to manifest the effect of selective enrichment of cell populations with phenotype of hemopoietic progenitors (CD34+CD38– cells). Comparing the content of CFUs, CFU-GM, HSCs and cells with CD34+CD38– phenotype has shown that cryopreservation regimen for FL cells was not optimal for BM. It has been demonstrated that varying cryopreservation conditions allowed not only to provide optimal preservation rate of hemopoietic progenitors derived from FL and BM with various initial functional status, but to control it purposefully. Probl Cryobiol Cryomed 2014; 24(2):118-131.
Highlights
The performed comparative analysis of structural and functional status of fetal liver (FL) hematopoietic stem cells (HSCs) of 14 gestation days and adult bone marrow (BM) of 4-month-old CBA/H mice demonstrated the differences of their initial qualitative and quantitative characteristics and different responses to the effect of cryopreservation factors
Cells of FL and BM with the concentration of 6.0×106 cells/ml and volume of 1.0 ml were frozen with programmable freezer UOP-6 produced by Special Designing and Technical Bureau with Experimental Unit (IPC&C of the NAS of Ukraine, Kharkov) in plastic ampoules (Nunc, Germany) by the following regimens: – Regimen 1 (R1) was the cooling from room temperature down to –40°C with the cooling rate of 1 deg/min with 5% dimethyl sulfoxide (DMSO) [26]; проблемы криобиологии и криомедицины problems of cryobiology and cryomedicine том/volume 24, No/issue 2, 2014 зиям клеток фетальной печени (ФП) или костного мозга (КМ) по каплям добавляли криоконсервирующий раствор в соотношении 1:1 при температуре 18°С в течение 2 мин
Our results indicate a different character of change in the content of CD34+CD38– cells in BM and FL
Summary
The performed comparative analysis of structural and functional status of fetal liver (FL) hematopoietic stem cells (HSCs) of 14 gestation days and adult bone marrow (BM) of 4-month-old CBA/H mice demonstrated the differences of their initial qualitative and quantitative characteristics and different responses to the effect of cryopreservation factors. To estimate structural and functional characteristics of HSCs derived from different sources there were used cytofluorimetry (content of CD34+СD38– cells), methods of colony formation in vivo (CFUs) and in vitro (CFU-GM). Comparing the content of CFUs, CFU-GM, HSCs and cells with CD34+CD38– phenotype has shown that cryopreservation regimen for FL cells was not optimal for BM. Со временем для лечения патологий различного генеза стало возможным применение СКК из других источников, например, фетальной печени (ФП), кордовой крови и т. With the course of time the treatment of pathologies of different genesis became possible by applying the HSCs from other sources, e. g. fetal liver (FL), cord blood etc. [7, 15], the necessity to create their stocks requires cryopreservation, being a mandatory stage for using such a tissue substrate in clinical practice [12, 26]
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