Study of the effects of stromal vascular fraction, cultured adipose-derived stem cells, and paracrine factors of a conditioned medium in the treatment of severe radiation injuries of rat skin

2019 ◽  
pp. 24-32
Author(s):  
В.Г. Лебедев ◽  
Ю.Б. Дешевой ◽  
А.А. Темнов ◽  
Т.А. Астрелина ◽  
К.А. Рогов ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Conditioned Medium ◽  
Stromal Vascular Fraction ◽  
Planar Geometry ◽  
X Rays ◽  
Radiation Injuries ◽  
Paracrine Factors ◽  
Wistar Kyoto ◽  
Radiation Ulcers

Цель - изучение влияния трансплантации стромально-васкулярной фракции (СВФ), культивированных мультипотентных мезенхимальных стволовых клеток (ММСК) жировой ткани и введения паракринных факторов кондиционной среды, полученных при культивировании ММСК, на течение тяжелых местных радиационных поражений кожи. Методика. Крыс инбредной линии Wistar-Kyoto облучали на рентгеновской установке в дозе 110 Гр, при мощности дозы 20,0 Гр/мин. Напряжение на трубке 30 кВ, ток 6,1 мА, фильтр Al толщиной 0,1 мм, площадь поля облучения составляла 8,5 см2. Трансплантацию клеток СВФ жировой ткани проводили двукратно в дозах 2,2 х 106 и 3,0 х 106 на 28-е и 35-е сут после облучения, соответственно. Культивированные ММСК вводили в дозах 2,8 × 106 и 3,4 × 106 на 28-е и 35-е сут после действия радиации. Паракринные факторы кондиционной среды вводили пятикратно с 1-х по 10-е сут после облучения, подкожно по 1,0 мл (общий белок 8 мг/мл). Тяжесть лучевого поражения кожи и эффекты терапии оценивали в динамике по клиническим проявлениям, с помощью планиметрии и патоморфологических методов. Результаты. Выбранный режим радиационного воздействия вызывал тяжелые лучевые поражения кожи с длительно (до 5 мес) незаживающими язвами. Площадь лучевых язв у крыс контрольной группы в период с 28-х до 84-х сут медленно уменьшалась от 2,73±0,17 см2 до 1,52±0,17 см2, а через 4 и 5 мес после облучения составляла 0,52±0,15 см2 и 0,05±0,04 см2, соответственно. Во всех опытных группах после 84-х сут отмечалась тенденция к снижению площади лучевых язв, а через 4 мес после облучения уменьшение площади язв и снижение тяжести поражения в опытных группах по сравнению с контролем были статистически значимыми (р < 0,05). У 70-80 % крыс в опытных группах через 4 мес после облучения наблюдалась полная эпителизация радиационных ран с образованием атрофического рубца, а в контрольной группе в этот период атрофические рубцы наблюдались лишь у 40% крыс. Заключение. Трансплантация СВФ жировой ткани или культивированных ММСК, а также введение паракринных факторов кондиционной среды способствуют усилению регенераторных процессов в пораженной ткани, и могут быть в равной степени эффективны при терапии тяжелых лучевых поражений кожи в эксперименте. Aim. To study effects of stromal-vascular fraction (SVF) transplantation, cultured multipotent mesenchymal stem cells (MMSC) of adipose tissue, and administration of paracrine factors of the conditioned MMSC culture medium on the course of severe, local radiation lesions of the skin. Methods. Wistar-Kyoto rats were irradiated with X-rays at a dose of 110 Gy and a dose rate of 20.0 Gy/min. The tube voltage was 30 kV, current - 6.1 mA, filter -0.1 mm thick A1, and irradiation field area - 8.5 cm2. Transplantation of adipose tissue SVF cells was performed twice at doses of 2.2 x 106 and 3.0 x 106 on the 28th and 35th days after irradiation, respectively. Cultured MMSC was administered at doses of 2.8 x 106 and 3,4 x 106 on the 28th and 35th days after irradiation. Paracrine factors of the conditioned medium were administered five times from the 1st to the 10th day after irradiation at a dose of 1.0 ml, s.c. (total protein, 8 mg/ml). Severity of radiation damage of the skin and effects of the therapy were followed up by clinical manifestations using planar geometry and pathomorphological methods. Results. Radiation exposure under the specified conditions caused severe radiation injuries of the skin with nonhealing ulcers that persisted for up to 5 months after irradiation. In control rats the area of radiation ulcers slowly decreased between the 28th and 84th days from 2.73 ± 0.17 cm2 to 1.52 ± 0.17 cm2, respectively, and was 0.52 ± 0.15 cm2 and 0.05 ± 0.04 cm2 at 4 and 5 months after irradiation. In all experimental groups, the healing was more intensive after the 84th day with a tendency to decrease the area of radiation ulcers; at 4 months after irradiation, statistically significant decreases in the area of ulcers and severity of lesions were observed in the experimental groups compared to the control (p <0.05). At four months after irradiation, 70-80% of rats in the treatment groups showed complete epithelialization of radiation wounds with formation of an atrophic scar whereas in the control group during the same period, atrophic scars were observed only in 40% of rats. Conclusion. Transplantation of adipose-derived SVF or cultured adipose tissue MMSC as well as administration of paracrine factors of the conditioned medium contributed to the enhancement of regenerative processes in the affected tissue and can be equally effective in the treatment of severe experimental radiation lesions of the skin.

The Influence of Mesenchymal Stem Cells of Adipose Tissue and Paracrine Factors of Conditioned Medium on the Healing of Radiation Ulcers in the Treatment of Severe Radiation Injuries of Skin in Rats

2021 ◽  
Vol 66 (2) ◽  
pp. 5-12
Author(s):  
V Lebedev ◽  
Yu. Deshevoy ◽  
A. Temnov ◽  
T. Astrelina ◽  
K. Rogov ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Conditioned Medium ◽  
Control Group ◽  
X Rays ◽  
Radiation Injuries ◽  
Paracrine Factors ◽  
Local Radiation ◽  
Radiation Ulcers

Purpose: Studying of the effects transplantation of cultured mesenchymal stem cells of adipose tissue (MMSC) and adipose-derived stromal vascular fraction (SVF), as well as the introduction of paracrine factors (PF) of conditioned medium in an isolated or combined application for severe local radiation skin lesions in the experiment. Material and methods: Rats of the inbred Wistar–Kyoto strain were irradiated to local X-rays exposure in the iliolumbar region of the back at a dose of 110 Gy. The transplantation of cultured MMSC was performed twice at doses of 2.1 × 106 and 2.6 × 106 on the 28th and 35th days after irradiation. Adipose-derived SVF was administered at the same time in doses of 3.2 × 106 and 2.8 × 106, respectively. PF were administered five times from the 1st to the 10th day after irradiation. The severity of radiation damage to the skin and the effects of therapy were evaluated in dynamics by clinical manifestations, using planimetry and histological methods. Results: Radiation exposure with these parameters caused severe radiation injuries of the skin with non-healing ulcers formed by the 21–25th day after irradiation. The area of radiation ulcers in rats of the control group in the period from the 26th to the 83rd day slowly decreased from 2.76 ± 0.12 cm2 to 1.85 ± 0.13 cm2. In 50 % of the animals in the control group, ulcers persisted for more than 4 months after irradiation. In rats of the experimental groups, more intensive healing and a decrease in the area of radiation ulcers was noted. With isolated administration of cultured MMSC or SVF, a statistically significant decrease in the area of ulcers compared with the control was observed on the 104–125th day, and with the introduction of PF on the 83rd day after irradiation, p <0.05. In the control group, by the118th day after irradiation, radiation ulcers healed only in 25 % of rats, and in the experimental groups with isolated administration of cultured MMSC, SVF and PF in 40–55 % of the rats showed complete epithelialization of wounds with the formation of an atrophic scar. Under the conditions of combined use of stem cells and conditioned medium factors, the number of animals with complete healing of radiation ulcers was 85–100 % by 118th days, p <0.05. Conclusion: Thus, transplantation of cultured MMSC of adipose tissue and adipose-derived SVF, as well as the introduction of PF of conditioned medium, can enhance the regeneration processes and stimulate skin regeneration, promoting earlier healing of chronic radiation ulcers in severe local radiation injuries. Moreover, with the combined introduction of PF and adipose-derived stem cell transplantation, the effectiveness of the healing of radiation ulcers was increases.

Transplantion of autologous cells of stromal vascular fraction of adipose tissue in severe local radiation injuries of skin caused by x-ray

2017 ◽  
Vol 62 (1) ◽  
pp. 5-11 ◽  
Author(s):  
Бушманов ◽  
Andrey Bushmanov ◽  
Самойлов ◽  
Alyeksandr Samoylov ◽  
Добрынина ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Clinical Manifestations ◽  
Stromal Vascular Fraction ◽  
Control Group ◽  
Tissue Cell ◽  
X Rays ◽  
Radiation Injuries ◽  
X Ray ◽  
Local Radiation ◽  
Radiation Ulcers

Purpose: To investigate the effectiveness of autologous cells of stromal vascular fraction of adipose tissue in severe local radiation skin injuries after the exposure of rats to X-rays. Material and methods: Experiments were performed on Wistar rats, weighing 200–230 g. Rats were exposed locally in iliolumbar region using X-ray machine LNC-268 (RAP 100-10) at a dose of 110 Gy (30 kV tube voltage, current 6.1 mA, thick Al filter 0.1 mm), dose rate: 17.34 Gy/min. Area of the irradiation field was 8.2–8.5 cm2. Transplantation of autologous cells of stromal vascular fraction (SVFC) of adipose tissue was carried out on 21st or 35th days after irradiation. SVFC isolation was performed by means of enzymatic treatment of adipose tissue. SVFC suspension was administered subcutaneously at a dose of 1×106 cells per injection around the radiation ulcers.The severity of radiation damage to the skin and the effects of cellular therapy were evaluated in the dynamics of clinical manifestations, with the help of plane geometry and pathomorphometry. Results: It was found that by the 17–25th day after irradiation radiation ulcers were formedon rat skin. In the control group of animalsulcers persisted throughout the observation period of more than 3 months. The area of ulcers was 1,87 ± 0,35 cm2 and 1.52 ± 0.24 cm2 at 83th and 90th days after irradiation, respectively. In animals of the experimental group, with autologous stromal vascular fraction of adipose tissue, was significant decrease in ulceration the area in comparison to control animals. In 80 % of the rats treated with SVFC on 21st day after exposure, to the 90th day after irradiation complete healing of ulcers occurred with the formation of atrophic scar at the site of radiation injuries. These clinical observations and planimetric were correlated with the results of histomorphometry. Conclusion: Transplantation autologous SVFC of adipose tissue contributes to accelerate the healing of radiation ulcers after local x-ray exposure in the experiment, indicating that the prospects of using adipose tissue cell products for the treatment of severe local radiation injuries.

Investigation of the Influence of the Conditioning Medium Factors Obtained During the Cultivation of Bone Marrow Mesenchymal Stem Cells on the Course of Severe Local Radiation Injuries of Skin in Rats

2018 ◽  
Vol 63 (1) ◽  
pp. 35-43 ◽  
Author(s):  
А. Темнов ◽  
A. Temnov ◽  
Т. Астрелина ◽  
T. Astrelina ◽  
К. Рогов ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Skin Lesion ◽  
Conditioned Medium ◽  
Radiation Injuries ◽  
Lesion Area ◽  
Paracrine Factors ◽  
Local Radiation ◽  
Radiation Ulcers

Purpose: Study of the effect of paracrine factors, produced by MMSC of bone marrow during the cultivation, on the severity of local radiation injuries in the conditions of application in the early periods after irradiation. Material and methods: Experiments were performed on rats of the breed Wistar weighing 280 g. Rats were exposed locally in iliolumbar region of the back using X-ray machine LNC-268 (RAP 100-10) at a dose of 110 Gy (30 kV tube voltage, current 6.1 mA, filter Al 0.1 mm thick), dose rate is 21.4 Gy/min. Area of the irradiation field was 8.2–8.5 cm2. The conditioned medium obtained by culturing MMSC of rats’ bone marrow was administered in dose 1.0 ml (total protein 8 mg/ml) at 1, 3, 6, 8 and 10 days after irradiation. The severity of radiation damage to the skin and the effects of therapy were evaluated in dynamics by clinical manifestations, using planimetry and histological methods. Results: It was shown that in control animals and in rats, with the introduction of the conditioned medium, the values of the skin lesion area in the period up to the 29th day after irradiation practically did not differ, gradually decreasing in control animals from 5.9 ± 0.6 cm2 to 2.2 ± 0.3 cm2 at the 15th and 29th days after irradiation, respectively. Then, in the control group, the lesion area ranged from 1.4 ± 0.6 cm2 on the 50th day to 1.9 ± 0.8 cm2 on the 71st day. In the experimental group of animals, with the introduction of factors of the conditioning medium, a decrease in the area of the lesion and a stable dynamics of healing of radiation ulcers, beginning from the 36th day, there was a gradual decrease in the area of the lesion, which reached 0.2 ± 0.1 cm2 by the 71st day after irradiation. On the 64–71th day after irradiation, the difference between the areas of skin lesion in the experimental and control groups was statistically significant, p <0.05. The histological analysis showed that the use of paracrine factors obtained from MMSC in the process of cultivation significantly reduces the severity of the inflammatory reaction and accelerates the regeneration processes. Conclusion: Thus, the introduction of conditioned medium factors obtained during the cultivation of mesenchymal stem cells of the bone marrow facilitates a more easy flow of the pathological process and the healing of radiation ulcers after local radiation damage to the skin of rats. Apparently, the favorable effect of paracrine factors introduced in the early periods after irradiation, with severe local radiation injuries, is associated with their effect on pathological processes in the inflammatory-destructive stage.

Effect of cultivated multipotent mesenchymal stromal cells of adipose tissue to restore skin with local radiation lesions

2017 ◽  
pp. 62-66
Author(s):  
Ю.Б. Дешевой ◽  
Б.Б. Мороз ◽  
Т.А. Насонова ◽  
В.Г. Лебедев ◽  
О.А. Добрынина ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Radiative Forcing ◽  
Clinical Manifestations ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Skin Lesions ◽  
Planar Geometry ◽  
Local Irradiation ◽  
Wistar Kyoto ◽  
Radiation Ulcers

Цель - изучение лечебной эффективности сингенных мезенхимальных стволовых клеток (ММСК) при тяжелых лучевых поражениях кожи. Методы. Эксперименты проводили на крысах инбредной линии Wistar-Kyoto. Локальное облучение в дозе 110 Гр (напряжение на трубке 30 кВ, ток 6,1 мА, фильтр Al толщиной 0,1 мм) выполняли на рентгеновской установке. Мощность дозы - 17,34 Гр/мин. Площадь облучения - 8,5 см. Такое радиационное воздействие позволяло получать тяжелые лучевые поражения кожи с длительно (до 3,5 мес.) не заживающими язвами. ММСК выделяли из подкожной жировой ткани интактных животных с помощью коллагеназы и затем культивировали их in vitro . ММСК вводили (1,6-1,8 х 10 клеток на одну инъекцию) под кожу вокруг лучевых язв. Тяжесть лучевого поражения кожи и эффекты клеточной терапии оценивали в динамике по клиническим проявлениям, с помощью планиметрии и патоморфологических методов. Результаты. Установлено, что введение культивированных сингенных ММСК дважды через 27 и 34 сут. или через 34 и 42 сут. после локального облучения, способствует усилению регенераторных процессов в пораженной ткани, что проявлялось более быстрым заживлением лучевых язв у леченых животных по сравнению с облученным контролем. Заключение. Полученные результаты свидетельствуют, что трансплантированные аутологичные ММСК, полученные из жировой ткани, могут быть эффективны при лечении длительно незаживающих лучевых язв. The purpose. Study of the therapeutic effectiveness of syngeneic mesenchymal stem cells (MMSK) with heavy radiation skin lesions. Methods. Experiments conducted on rats Wistar-Kyoto inbreed. Local irradiation dose of 110 Gy (voltage on the tube 30 kV, current 6.1 Ma, filter 0.1 mm thick Al) served through an x-ray machine. Dose rate is 17.34 Gy/min. Area field irradiation was 8.5 cm. Radiative forcing is allowed to receive severe radiation damage of the skin with long (up to 3.5 months) did not heal ulcers. MMSK singled out of the subcutaneous adipose tissue intact animals using collagenase and then cultivated in vitro . MMSK injected (1.6-1.8 х 10 cells per injection) under the skin around the radiation ulcers. The severity of radiation injury to the skin and the effects of cell therapy were evaluated in the dynamics of clinical manifestations, using planar geometry and pathomorphological methods. Results. It has been established that the introduction of cultivated syngeneic MMSK, investigated through 27 and 34 days or twice through 34 and 42 days after local irradiation enhances regenerative processes in the affected tissue, which showed faster healing Ray treated ulcers in animals compared to the irradiated control. Conclusion. The results showed that the transplanted autologous MMSK derived from adipose tissue, can be effective in the treatment of long-term healing of radiation ulcers.

scholarly journals Application of Regenerative Cell Technologies in the Treatment of Severe Local Radiation Injuries in Victims of Emergency Situations: from the Experience of the Specialists of the Federal Medical Biophysical Center Named after A.I. Burnazyan of the Federal Medical and Biological Agency of Russia

2021 ◽  
pp. 58-64
Author(s):  
T.A. Astrelina ◽  
◽  
A.V. Aksenenko ◽  
I.V. Kobzeva ◽  
V.A. Brunchukov ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Stromal Vascular Fraction ◽  
Healing Time ◽  
Radiation Injuries ◽  
Serious Adverse Events ◽  
Emergency Situations ◽  
Satisfactory Condition ◽  
Complex Therapy ◽  
Local Radiation ◽  
Radiation Ulcers

It has been noted that one of the most common types of radiation injuries when a person is exposed to ionizing radiation is radiation burns — severe local radiation injuries. The aim of the study is to apply stromal and vascular fraction of adipose tissue to increase the efficiency of complex therapy for local radiation lesions. Materials and methods of the study. In 2017-2019, 7 patients (all male; mean age — (54.83±9.41) years) with local radiation lesions of the skin — ulcerative-necrotic lesions of the skin and underlying tissues — were treated at the Federal Medical Biophysical Center named after A.I. Burnazyan of FMBA of Russia. For more than 6 months the patients received conventional conservative therapy of local radiation lesions and a single injection of cell suspension of autologous cells of stromal-vascular fraction of adipose tissue — the average number of cells was (60.33×106±64.04). Results of the study and their analysis. All patients had no serious adverse events and reactions associated with the introduction of autologous regenerative cells of adipose tissue. During the whole period of observation after stromal-vascular fraction of adipose tissue was injected, late radiation ulcers remained without signs of inflammation and infiltration. The patients were discharged from the hospital in satisfactory condition. According to the authors, the use of stromal-vascular fraction of adipose tissue in local radiation lesions provides favorable conditions: to increase the effectiveness of complex therapy; to reduce healing time of the wound surface; to regulate and activate immune and reparative processes in the dermis; to restore the damaged vascular network, lost skin without severe scarring changes; to heal and achieve a satisfactory result, decent quality of life of patients.

scholarly journals Derivation, characterization, and in vitro cell regeneration of canine white adipose tissue-derived mesenchymal stem cells obtained from a mesenteric region

2021 ◽  
Vol 82 (1) ◽  
Author(s):  
Anirban Mandal ◽  
Ajeet Kumar Jha ◽  
Dew Biswas ◽  
Shyamal Kanti Guha
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
White Adipose Tissue ◽  
Stromal Vascular Fraction ◽  
Cell Regeneration ◽  
Wound Healing Assay ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background The study was conducted to assess the characterization, differentiation, and in vitro cell regeneration potential of canine mesenteric white adipose tissue-derived mesenchymal stem cells (AD-MSCs). The tissue was harvested through surgical incision and digested with collagenase to obtain a stromal vascular fraction. Mesenchymal stem cells isolated from the stromal vascular fraction were characterized through flow cytometry and reverse transcription-polymerase chain reaction. Assessment of cell viability, in vitro cell regeneration, and cell senescence were carried out through MTT assay, wound healing assay, and β-galactosidase assay, respectively. To ascertain the trilineage differentiation potential, MSCs were stained with alizarin red for osteocytes, alcian blue for chondrocytes, and oil o red for adipocytes. In addition, differentiated cells were characterized through a reverse transcription-polymerase chain reaction. Results We observed the elongated, spindle-shaped, and fibroblast-like appearance of cells after 72 h of initial culture. Flow cytometry results showed positive expression for CD44, CD90, and negative expression for CD45 surface markers. Population doubling time was found 18–24 h for up to the fourth passage and 30±0.5 h for the fifth passage. A wound-healing assay was used to determine cell migration rate which was found 136.9 ± 4.7 μm/h. We observed long-term in vitro cell proliferation resulted in MSC senescence. Furthermore, we also found that the isolated cells were capable of differentiating into osteogenic, chondrogenic, and adipogenic lineages. Conclusions Mesenteric white adipose tissue was found to be a potential source for isolation, characterization, and differentiation of MSCs. This study might be helpful for resolving the problems regarding the paucity of information concerning the basic biology of stem cells. The large-scale use of AD-MSCs might be a remedial measure in regenerative medicine.

Eine aktuelle Übersicht über die Einflussfaktoren der Stammzellspender auf das regenerative Potential von Fettgewebsstammzellen

2020 ◽  
Vol 52 (06) ◽  
pp. 521-532
Author(s):  
Constanze Kuhlmann ◽  
Thilo Ludwig Schenck ◽  
Elisabeth Maria Haas ◽  
Riccardo Giunta ◽  
Paul Severin Wiggenhauser
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Stromal Vascular Fraction ◽  
National Library ◽  
Regenerative Therapie

Zusammenfassung Hintergrund Nicht nur regenerative Therapie wie zellassistierter Lipotransfer (cell assisted lipotransfer) sondern auch präklinische experimentelle Studien verwenden in der Plastischen Chirurgie Stammzellen aus Fettgewebe, sogenannte Adipose tissue-derived Stem Cells (ASCs). Hierbei haben allerdings vom jeweiligen Stammzellspender abhängige Faktoren einen entscheidenden Einfluss auf die Zellausbeute und das regenerative Potential von ASCs und der Stromal vascular Fraction (SVF). Ziel dieser Übersichtsarbeit war es daher, diese Einflussfaktoren des Stammzellspenders darzustellen und anhand des aktuellen Wissenstands zu beurteilen. Methoden Es erfolgte eine intensive Literaturrecherche in der der National Library of Medicine, mit Fokus auf Einflussfaktoren der Stammzellspender, die eine Beeinflussung der Zellausbeute und des regenerativen Potentials von humanen ASCs und SVF in vorherigen Studien gezeigt haben. Ergebnisse Aktuell gibt es eine Vielzahl von Studien, welche sich mit den Einflussfaktoren des Stammzellspenders auseinandersetzen. Allerdings sind diese Faktoren sehr inhomogen und teilweise sogar widersprüchlich, so dass hier noch weiterer Forschungsbedarf besteht. Dennoch gibt es einige Faktoren, die gemäß der aktuellen Literatur gehäuft untersucht wurden: Alter, Geschlecht, Gewicht, Nebenerkrankungen (z. B. Diabetes, Lipödem) sowie spezielle Medikamente (Antidepressiva, Antihormontherapie) und Chemotherapie. Schlussfolgerung Wir empfehlen, bei experimentellen und klinischen Arbeiten mit ASCs/SVF eine Charakterisierung des Patientenkollektivs zu veröffentlichen, so dass mögliche Beeinflussungen durch oben genannte Faktoren kommuniziert werden und eine bessere Vergleichbarkeit von Studien ermöglicht wird. Darüber hinaus kann aber auch mit einer präzisen Anamnese und körperlichen Untersuchung vorab ein möglichst homogenes Patientenkollektiv für die Sammlung von Proben für wissenschaftliche Arbeiten konstruiert werden. Auch könnten die Ergebnisse dazu beitragen, den Erfolg zukünftiger ASC-basierter Therapien einzuschätzen.

scholarly journals Adipose Derived-Mesenchymal Stem Cells Viability and Differentiating Features for Orthopaedic Reparative Applications: Banking of Adipose Tissue

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Ilaria Roato ◽  
Daniela Alotto ◽  
Dimas Carolina Belisario ◽  
Stefania Casarin ◽  
Mara Fumagalli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Articular Cartilage ◽  
Tissue Banking ◽  
Stromal Vascular Fraction ◽  
Single Infusion ◽  
Knee Arthritis ◽  
Symptomatic Knee

Osteoarthritis is characterized by loss of articular cartilage also due to reduced chondrogenic activity of mesenchymal stem cells (MSCs) from patients. Adipose tissue is an attractive source of MSCs (ATD-MSCs), representing an effective tool for reparative medicine, particularly for treatment of osteoarthritis, due to their chondrogenic and osteogenic differentiation capability. The treatment of symptomatic knee arthritis with ATD-MSCs proved effective with a single infusion, but multiple infusions could be also more efficacious. Here we studied some crucial aspects of adipose tissue banking procedures, evaluating ATD-MSCs viability, and differentiation capability after cryopreservation, to guarantee the quality of the tissue for multiple infusions. We reported that the presence of local anesthetic during lipoaspiration negatively affects cell viability of cryopreserved adipose tissue and cell growth of ATD-MSCs in culture. We observed that DMSO guarantees a faster growth of ATD-MSCs in culture than trehalose. At last, ATD-MSCs derived from fresh and cryopreserved samples at −80°C and −196°C showed viability and differentiation ability comparable to fresh samples. These data indicate that cryopreservation of adipose tissue at −80°C and −196°C is equivalent and preserves the content of ATD-MSCs in Stromal Vascular Fraction (SVF), guaranteeing the differentiation ability of ATD-MSCs.

scholarly journals Adipose tissue derived stromal vascular fraction transplantation can recover spinal cord injury in mice

2016 ◽  
Vol 3 (04) ◽  
pp. 144 ◽  
Author(s):  
Hau Thi-My Lam ◽  
Minh Nguyen-Thu Tran ◽  
Khoa Anh Bui ◽  
Thao Thi-Thu Le ◽  
Khanh Hong-Thien Bui ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Adipose Tissue ◽  
Mouse Model ◽  
Platelet Rich Plasma ◽  
Stromal Vascular Fraction ◽  
Paracrine Factors ◽  
Diabetic Ulcer ◽  
Motor Test ◽  
Cord Injury

Introduction: Stem cell therapy is one of the most promising therapies for degenerative diseases and related injuries. Adipose tissue derived stem cells (ADSCs) exhibit some particular properties such as high production of paracrine factors. Indeed, ADSCs have been successfully used to treat diseases, including osteoarthritis, diabetic ulcer, etc. Methods: In this study, ADSCs were used to treat spinal cord injury (SCI) in a mouse model. Non-expanded ADSCs, from stromal vascular fractions (SVFs) isolated from both autologous and allogeneic adipose tissues, were injected into injured sites of mice at a specified dose. The SCI mouse model were generated by transection of spinal cord at vertebrae T8 - T10. After 1 week of transection, mice exhibiting completed SCI were divided into 4 groups: group 1 was control (mice without any treatment), group 2 was placebo (mice treated with platelet rich plasma (PRP)), group 3 was allogeneic SVF transplantation (mice treated with allogeneic SVFs), and group 4 was autologous SVF transplantation (mice treated with autologous SVFs). For the treatment groups, mice were transplanted with 20 µL of activated PRP or/and with 106 cells of SVF (allogeneic or autologous) into the injured position through laminectomy. The recovery of SCI was evaluated by locomotor test, sensory test and sensory-motor test at 5 weeks after transplantation. The histology of the spinal cord also was checked after 5 weeks. Results: The results showed that in all groups with PRP injected with or without SVFs, the inflammation was efficiently controlled. The glial scar as well as myelin defragmentation were clearly reduced. However, a significant improvement of BBB score was only recorded in mice transplanted with autologous SVFs. Conclusion: The results of our study show that autologous SVF transplantation in combination with PRP can be a promising therapy for SCI.

Abstract 362: Proteomic Analysis of Low Oxygen Tension-Induced Secretome of Adipose Tissue-Derived Stem Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Zeljko Bosnjak ◽  
Bassam Wakim ◽  
Yasheng Yan ◽  
Scott Canfield ◽  
Chika Kikuchi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Oxygen Tension ◽  
Proteomic Analysis ◽  
Stem Cell Markers ◽  
Paracrine Factors ◽  
Differentiation Potential ◽  
Trophic Effect ◽  
Lower Oxygen

Growing evidence from animal studies shows that adipose tissue-derived stem cells (ASCs) improve cardiac function of infarcted hearts. It is commonly accepted that therapeutic potential of ASCs may depend more on their paracrine effects than differentiation potential. The underlying mechanisms remain unclear. However, most data regarding paracrine factors were obtained from ASCs cultured in normoxic condition (20%). The present study investigated how in vivo physiological oxygen (4%) tension influenced the secretome of ASCs. ASCs were isolated from three 8-week-old BALB/c mice. ASCs were confirmed by the expression of stem cell markers (CD44 and CD90) and their capacity to differentiate into adipocytes and osteocytes. ASCs at passage 5 were cultured in normoxic (20%) and lower oxygen (4%) incubators and conditioned for 24 h (3 cultures/group). The conditioned media (CM) from ASCs were subjected to trypsin digestion followed by analysis using automated nano-flow liquid chromatography tandem mass spectrometry. The collected LC/MS/MS data were searched against the rodent subset of the Uniprot database and the total proteomes were identified. The data were from 6 technical replicates. A total of 28 proteins were identified and 7 proteins were unique to normoxic CM. Of the 21 common proteins detected in both normoxic and lower oxygen CM, 9 were extracellular matrix proteins. The abundance of 6 of these proteins (e.g., collagen I and laminin) differed noticeably between normoxic and lower oxygen CM. In addition, a greater amount of cytokine CXCL5 and matrix metalloproteinase (MMP)-2 was detected in lower oxygen CM than in normoxic CM while tissue inhibitor of metalloproteinase (TIMP)-1 was only detected in normoxic CM. These results indicate that lower oxygen tension differentially regulates the secretome of ASCs. Extrapolating the results of this study to the in vivo setting, it would appear that injected ASCs may exert their anti-fibrotic and trophic effect by 1) directly regulating the balance of MMP/TIMP production and preventing collagen accumulation in ischemic hearts to decrease fibrosis, and 2) secreting trophic factors including CXCL5. These data suggest that proteomic analysis of CM is useful for elucidation of the paracrine effect of ASCs in vivo.

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