scholarly journals Possibility of using combined transplantation of multipotent mesenchymal stromal cells and hepatic stellate cells to activate reparative liver regeneration in mature and old organism

2020 ◽  
Vol 22 ◽  
pp. 01004
Author(s):  
Irina Maklakova ◽  
Dmitry Grebnev ◽  
Victoria Vakhrusheva ◽  
Ekaterina Petrunina
Keyword(s):  
Body Weight ◽  
Liver Regeneration ◽  
Cell Transplantation ◽  
Stromal Cells ◽  
Age Groups ◽  
Stellate Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Laboratory Animals ◽  
Subtotal Resection ◽  
Intracellular Regeneration

The aim of the study was to examine the effect of combined transplantation of multipotent mesenchymal stromal (MMSC) and hepatic stellate (HSC) cells on liver regeneration after resection. Research has been carried out on laboratory animals of mature and old age. After the subtotal resection of the liver, MMSC and HSC were introduced in the tail vein in the amount of 4 million cl/kg of body weight and 9 million cl/kg of body weight respectively. Evaluation of reparative liver regeneration was performed on the 1st, 3rd, 7th days after subtotal liver resection. Features of reparative liver regeneration in mature and old organism were revealed. In mature organism against the background of combined cell transplantation, regeneration activation is achieved by increasing cellular and intracellular regeneration mechanisms. In this case, the old organism responds to cell transplantation by activating only intracellular mechanisms. In both age groups, decreased mutagenesis and inhibition of programmed cell death against the background of MMSC cotransplantation and HSC were observed.

THE EFFECT OF MULTIPOTENT MESENCHYMAL STROMAL CELLS TRANSPLANTATION ON LIVER MORPHOMETRIC PARAMETERS OF MATURE AND OLD LABORATORY ANIMALS WITH TOXIC HEPATITIS

2020 ◽  
pp. 55-60
Author(s):  
И. Ю. Маклакова ◽  
Д. Ю. Гребнев ◽  
В. Ч. Юсупова ◽  
Е. М. Петрунина
Keyword(s):  
Mitotic Activity ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Toxic Hepatitis ◽  
Age Groups ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Morphometric Parameters ◽  
Laboratory Animals ◽  
Intracellular Regeneration ◽  
Hepatocyte Nucleus

Цель - изучение влияния трансплантации мультипотентных мезенхимальных стромальных клеток (ММСК) на морфометрические показатели печени зрелых и старых лабораторных животных в условиях токсического гепатита. Материал и методы. Эксперименты выполнены на зрелых и старых мышах-самцах. Токсический гепатит вызывали путем внутрибрюшинного введения CClв дозе 50 мкг/кг. Трансплантация клеток осуществлялась в хвостовую вену через 1 ч после введения четыреххлористого углерода однократно. Исследовалось влияние ММСК на морфометрические показатели печени в физиологических условиях и условиях токсического гепатита на 1-, 3-, 7-е сутки после трансплантации клеток. Результаты. У зрелых лабораторных животных на 3-и сутки после введения ММСК на фоне токсического гепатита обнаружено увеличение митотической активности, повышение количества гепатоцитов, площади ядра гепатоцитов и ядерноцитоплазматического индекса. В то же время, у старых лабораторных животных выявлено лишь увеличение площади ядра гепатоцитов и ядерно-цитоплазматического индекса. На 7-е сутки после введения ММСК на фоне токсического гепатита в обеих возрастных группах выявлены активация митотической активности, повышение количества гепатоцитов, увеличение площади ядра гепатоцитов и ядерно-цитоплазматического индекса. Выводы. Изменение морфометрических показателей печени у зрелых и старых лабораторных животных реализуется через механизмы как клеточной, так и внутриклеточной регенерации. При этом у старых лабораторных животных на 3-и сутки после введения ММСК выявлена активация лишь внутриклеточной регенерации, в то время как у зрелых лабораторных животных имеет место повышение клеточной и внутриклеточной регенерации гепатоцитов. В более поздние сроки в обеих изучаемых возрастных группах изменение основных морфометрических показателей печени реализуется через активацию как клеточной, так и внутриклеточной регенерации. Objective - to study the influence of multipotent mesenchymal stromal cells (MMSC) transplantation on morphometric parameters of the liver of mature and old laboratory animals with toxic hepatitis. Material and methods. The experiments were performed on mature and old male mice. Toxic hepatitis was caused by intraperitoneal administration of CCl4 at a dose of 50 μg/kg. The cells were transplanted via the tail vein 1 hour after administration of a single dose of carbon tetrachloride. The effect of MMSC on liver morphometric parameters in physiological conditions and after toxic hepatitis development was studied on days 1, 3, 7 after cell transplantation. Results. An increase in mitotic activity, an increase in the number of hepatocytes, hepatocyte nucleus area, and nuclear cytoplasmic index were found in mature laboratory animals with toxic hepatitis on the 3 day after the introduction of MMSC. At the same time, only an increase in the area of hepatocyte nucleus and nuclear cytoplasmic index was revealed in old laboratory animals. On the 7 day after the introduction of MMSC to the animals with toxic hepatitis, both age groups demonstrated activation of mitotic activity, an increase in the number of hepatocytes, an increase in the area of hepatocyte nucleus and nuclear cytoplasmic index. Conclusions. Changes in liver morphometric parameters in mature and old laboratory animals are realized through mechanisms of both cellular and intracellular regeneration. In addition, the activation of only intracellular regeneration was found in old laboratory animals on the 3rd day after the introduction of MMSC, while in mature laboratory animals there was an increase in cellular and intracellular regeneration of hepatocytes. In later periods in both studied age groups, the change in the main liver morphometric parameters is realized through the activation of both cellular and intracellular regeneration.

scholarly journals Activation of reparative liver regeneration following the combined transplantation of multipotent mesenchymal stromal cells and hepatic stellate cells

2021 ◽  
Vol 102 (5) ◽  
pp. 669-677
Author(s):  
I Yu Maklakova ◽  
D Yu Grebnev ◽  
A V Osipenko
Keyword(s):  
Liver Regeneration ◽  
Partial Hepatectomy ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Hepatic Stellate Cells ◽  
Comparison Group ◽  
Stellate Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Control Group ◽  
Nacl Solution

Aim. To study the effect of combined transplantation of multipotent mesenchymal stromal and hepatic stellate cells on the reparative liver regeneration. Methods. Laboratory mice were given intravenous administration of multipotent mesenchymal stromal and hepatic stellate cells after partial hepatectomy. The mice were divided into four groups: control, experimental 1 (injection of multipotent mesenchymal stromal cells), experimental 2 (co-transplantation of multipotent mesenchymal stromal cells and hepatic stellate cells), the comparison group. Comparison of the experimental groups with the control group and the comparison group was carried out. Each group consisted of 14 animals. The control and experimental groups underwent partial hepatectomy. The experimental mice were injected with the cells into the lateral tail vein 1 hour after the operation. Multipotent mesenchymal stromal cells were administered at a dose of 4 million cells/kg (120 thousand cells/mouse), hepatic stellate cells in the amount of 9 million cells/kg (270 thousand cells/mouse), suspended in 0.2 ml 0.9% NaCl solution. The control group animals were injected with 0.2 ml 0.9% NaCl solution into the lateral tail vein. The comparison group consisted of mice without partial hepatectomy, injected with 0.2 ml 0.9% NaCl solution. To assess reparative regeneration of the liver, morphometric parameters of the liver, blood biochemical parameters on the 3rd and 7th days after partial hepatectomy were studied. The severity of apoptosis was assessed by the immunohistochemical method, the activity of deoxyribonucleic acid (DNA) repair enzymes of the poly (ADP-ribose) polymerases was determined by flow cytometry. The number of micronucleated hepatocytes was also determined. The hepatocyte growth factor (HGF) content was measured by using an enzyme-linked immunosorbent assay in serum. The significance of differences in the compared samples was determined by using the Student's t-test. Statistical data processing was performed by using the SPSS Statistics software version 17.0. Results. It was found that the combined transplantation of multipotent mesenchymal stromal and stellate liver cells causes restoration of the activity of alanine aminotransferase (a decrease of 30.3%, p=0.016), aspartate aminotransferase (a decrease of 27.7%, p=0.021), alkaline phosphatase (a decrease of 21.1%, p=0.036), an increase in the protein synthetic function of the liver (increase in albumin level by 36.6%, p=0.009), an increase in hepatocyte growth factor level by 74.3%. These changes were accompanied by the restoration of liver morphometric parameters: there was an increase in the mitotic activity of hepatocytes by 28.7% (p=0.008), the nuclear area of hepatocytes by 26.7% (p=0.006), the number of binucleated hepatocytes by 26.1% (p=0.004), which led to the restoration of liver mass. There was a decrease in the level of apoptosis by 28.8% (p=0.006) and a decrease in the number of micronucleated hepatocytes by 22.7% (p=0.001) compared with the control group, which may be related to an increase in the activity of Poly (ADP-ribose) polymerase repair enzymes detected in the study. The deviations were presented as a difference relative to the indicators of the control group (operated animals that were injected with 0.9% NaCl solution). Conclusion. Combined transplantation of multipotent mesenchymal stromal and hepatic stellate cells activates reparative liver regeneration after partial hepatectomy.Keywords: multipotent mesenchymal stromal cells, MSC, hepatic stellate cells, HSC, liver regeneration, partial hepatectomy.

EFFECT OF COMBINED TRANSPLANTATION OF MULTIPOTENT MESENCHYMAL STROMAL AND HEMATOPOIETIC STEM CELLS ON LIVER REGENERATION AFTER LIVER RESECTION IN OLD LABORATORY ANIMALS

2020 ◽  
Vol 17 (2) ◽  
pp. 139-148
Author(s):  
I.Yu. Maklakova ◽  
◽  
V.V. Bazarniy ◽  
D.Yu. Grebnev ◽  
◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Liver Regeneration ◽  
Stromal Cells ◽  
Morphometric Parameters ◽  
Laboratory Animals ◽  
Subtotal Resection ◽  
Laboratory Mice ◽  
Hematopoietic Stem ◽  
Cellular Regeneration

The aim of this study was to study the changes in the morphological and functional state of the liver of old laboratory animals after its resection against the background of combined transplantation of multipotentmesenchymal stromal and hematopoietic stem cells.Materials and methods. The studies were performed on old laboratory animals, male mice of the age of 16-17 months. Resection of 2/3 of the liver was performed in old laboratory mice according to the method of C. Mitchell and H. Willenbring. One hour after subtotal resection, a combined transplantation of two types of cells was performed: multipotentmesenchymal stromal cells (at a dose of 4 million cells / kg) and hematopoietic stem cells (at a dose of 330 thousand cells / kg). The experiments on obtaining the MMSC and HSC culture were performed from the placenta chorion of 10 laboratory female mice 3-4 months old, weighing 20-25 g, gestational age 18 days. Assessment of liver regeneration was carried out on days 1, 3, 7 after resection of the liver by analyzing morphometric parameters of the liver and biochemical parameters of peripheral blood.Results. Studies have shown the activation of protein synthesizing liver function in old animals, a decrease in the level of cytolysis enzymes, an increase in the activity of cellular regeneration (increase in the hepatocyte nucleus area, the number of binuclear cells) of liver regeneration after resection against the introduction of stem cells.

scholarly journals Homing and Engraftment of Intravenously Administered Equine Cord Blood-Derived Multipotent Mesenchymal Stromal Cells to Surgically Created Cutaneous Wound in Horses: A Pilot Project

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1162
Author(s):  
Suzanne J. K. Mund ◽  
Eiko Kawamura ◽  
Awang Hazmi Awang-Junaidi ◽  
John Campbell ◽  
Bruce Wobeser ◽  
...  
Keyword(s):  
Cord Blood ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Adverse Reactions ◽  
Pilot Project ◽  
Multipotent Mesenchymal Stromal Cells ◽  
The Body ◽  
Laboratory Animals ◽  
Clinical Safety ◽  
Allogeneic Mscs

Limb wounds on horses are often slow to heal and are prone to developing exuberant granulation tissue (EGT) and close primarily through epithelialization, which results in a cosmetically inferior and non-durable repair. In contrast, wounds on the body heal rapidly and primarily through contraction and rarely develop EGT. Intravenous (IV) multipotent mesenchymal stromal cells (MSCs) are promising. They home and engraft to cutaneous wounds and promote healing in laboratory animals, but this has not been demonstrated in horses. Furthermore, the clinical safety of administering >1.00 × 108 allogeneic MSCs IV to a horse has not been determined. A proof-of-principle pilot project was performed with two horses that were administered 1.02 × 108 fluorescently labeled allogeneic cord blood-derived MSCs (CB-MSCs) following wound creation on the forelimb and thorax. Wounds and contralateral non-wounded skin were sequentially biopsied on days 0, 1, 2, 7, 14, and 33 and evaluated with confocal microscopy to determine presence of homing and engraftment. Results confirmed preferential homing and engraftment to wounds with persistence of CB-MSCs at 33 days following wound creation, without clinically adverse reactions to the infusion. The absence of overt adverse reactions allows further studies to determine effects of IV CB-MSCs on equine wound healing.

THE STUDY OF MMSC HOMING AFTER LIVER RESECTION

2019 ◽  
pp. 40-45
Author(s):  
И.Ю. Маклакова ◽  
Д.Ю. Гребнев ◽  
В.Ч. Юсупова ◽  
Е.А. Примакова
Keyword(s):  
Liver Resection ◽  
Portal Vein ◽  
Hepatic Artery ◽  
Stromal Cells ◽  
Peripheral Blood ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Cell Number ◽  
Laboratory Animals ◽  
Cell Injection ◽  
Physiological Conditions

Цель работы - изучение направленного движения мультипотентных мезенхимальных стромальных клеток (ММСК), выделенных из плаценты, в различные органы и ткани лабораторных животных в физиологических условиях и после субтотальной резекции печени. Методика. Клеточную культуру ММСК получали из хориона плаценты лабораторных крыс. Культивирование ММСК проводилось в условиях СО2- инкубатора при температуре 37 ºC с содержанием углекислого газа 5% и влажностью 90%. Замену среды проводили каждые 3-4 сут до достижения клетками 70-80 % конфлюэнтности. При формировании соответствующего монослоя осуществлялся пересев клеток. При трансплантации лабораторным животным была использована культура ММСК 3-го пассажа. Введение клеток производили сразу после выполнения субтотальной резекции печени. Резекция 70% печени у крыс выполнена по методике G.M. Higgins, R.M. Anderson. Введение ММСК, меченых акридиновым оранжевым, осуществляли внутривенно, интраперитонеально, в печеночную артерию, в портальную вену в дозе 4 млн кл/кг. массы тела. Анализ распределения ММСК проводили через 3 и 24 ч. Результаты. Показано, что через 1 сут после введения клеток ложно-оперированным животным не отмечено существенных изменений распределения ММСК по сравнению с их распределением через 3 ч. Однако если введение клеток сопряжено с оперативным вмешательством (лапаротомия для обеспечения введения клеток в v. portae и a. hepatica) происходит снижение их количества в периферической крови. Через 1 сут после резекции печени в изучаемых органах и тканях (периферическая кровь, легкое, селезенка, костный мозг, тонкий кишечник, почка) содержание трансплантированных ММСК ниже по сравнению с их количеством в тот же период времени в данных органах и тканях без резекции печени. Обращает внимание значительное увеличение количества введенных клеток в печени после ее резекции как через 3 ч, так и через 24 ч по сравнению с физиологическими условиями. Заключение. Отмечено значительное увеличение количества клеток в печени после ее резекции вне зависимости от способа введения. Внутрибрюшинный способ введения клеток показал низкую эффективность доставки клеток в органы и ткани организма. The aim was to study migration of multipotent mesenchymal stromal cells (MSCS) isolated from the placenta to various organs and tissues of laboratory animals under the physiological conditions and after subtotal liver resection. Method. The MMSC cells for culturing were obtained from the rat placental chorion. MMSCs were cultured in a CO2 incubator at 37 oC, 5% CO2, and 90% humidity. The medium was replaced every 3-4 days until the cells reached 70-80 % confluence. Upon formation of an appropriate monolayer the cells were passed. The third passage culture of MMSC was used for transplantation to laboratory animals. Cells were injected immediately after subtotal liver resection. The 70% rat liver resection was performed according to the method of G.M. Higgins and R.M. Anderson. The acridine orange labeled MMSC were injected intravenously, intraperitoneally, into the hepatic artery, and into the portal vein at a dose of 4x106 cells/kg body weight. The MMSC distribution was analyzed at 3 and 24 hours. Results. In 24 h after the cell injection in the absence of liver resection, no significant changes were observed in the MMSCs distribution compared to their distribution in 3 hours. However, when the cell injection was associated with a surgery (laparotomy to ensure the cell injection into the portal vein and hepatic artery) the cell number was decreased in the peripheral blood. At one day after liver resection, the content of transplanted MMSCs was lower in the studied organs and tissues (peripheral blood, lung, spleen, bone marrow, small intestine, and kidney) compared to the respective values without liver resection in the same period. Noticeably, the number of injected cells was significantly increased in the liver at both 3 and 24 hours after resection compared to the physiological conditions. Conclusion. The number of cells was significantly increased in the liver after resection regardless of the cell administration route. Intraperitoneal cell injection showed a low effectiveness of cell delivery to organs and tissues.

scholarly journals CYP2C19 phenotype and body-weight-guided voriconazole initial dose in infants and children after hematopoietic cell transplantation

2021 ◽  
Author(s):  
Takuto Takahashi ◽  
Maryam A. Mohamud ◽  
Angela R. Smith ◽  
Pamala A. Jacobson ◽  
Mutaz M. Jaber ◽  
...  
Keyword(s):  
Body Weight ◽  
Cell Transplantation ◽  
Hematopoietic Cell Transplantation ◽  
Age Groups ◽  
Model Performance ◽  
Therapeutic Index ◽  
Hematopoietic Cell ◽  
Therapeutic Drug ◽  
Dosing Regimens ◽  
Trough Concentrations

Background : Prophylactic voriconazole use is recommended in children undergoing hematopoietic cell transplantation (HCT). Dosing considerations are essential due to its narrow therapeutic index. Known covariates do not sufficiently explain large interindividual pharmacokinetic (PK) variability of voriconazole. Moreover, knowledge of voriconazole PK for age <2 years is limited. Objectives : We investigated genetic and clinical covariate association with voriconazole interindividual PK variability and subsequently simulated dosing regimens in children. Methods : This study was conducted as a part of a single-institution, phase I study of intravenous voriconazole in children undergoing HCT. We conducted a population PK analysis and tested covariate effects on voriconazole PK, including 67 genetic variants and clinical variables. Results : We analyzed plasma voriconazole and n-oxide metabolite concentrations from 58 children aged <21 years (n=12 in age <2 years). A two-compartment parent mixed linear/nonlinear model best described our data. CYP2C19 phenotype and body weight were significant covariates (both p<0.05). Our model performance in age <2 years was comparable to other age groups. Simulation of the final model suggested the following dosages to attain target steady-state trough concentrations of 1.5 - 5.0 mg/L in CYP2C19 normal phenotype: 16 mg/kg (weight <15 kg), 12 mg/kg (weight 15-30 kg), 10 mg/kg (weight >30 kg), whereas dosages were 33-50% lower for CYP2C19 poor/intermediate and 25-50% higher for CYP2C19 rapid/ultrarapid phenotypes. Conclusions : We propose a new starting dosage regimen, combined with therapeutic drug monitoring for intravenous voriconazole in children of all ages. Future studies should validate this dosing regimen.

scholarly journals No Contribution of Multipotent Mesenchymal Stromal Cells to Liver Regeneration in a Rat Model of Prolonged Hepatic Injury

Stem Cells ◽  
2006 ◽  
Vol 25 (3) ◽  
pp. 639-645 ◽  
Author(s):  
Felix C. Popp ◽  
Przemyslaw Slowik ◽  
Elke Eggenhofer ◽  
Phillip Renner ◽  
Sven A. Lang ◽  
...  
Keyword(s):  
Liver Regeneration ◽  
Mesenchymal Stromal Cells ◽  
Rat Model ◽  
Stromal Cells ◽  
Hepatic Injury ◽  
Multipotent Mesenchymal Stromal Cells
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