Desiccation Tolerance of Adult Stem Cells in the Presence of Trehalose and Glycerol

Development of protocols for storing desiccated cells at ambient temperatures offers tremendous economic and practical advantages over traditional storage procedures like cryopreservation and freeze-drying. As a first step for developing such procedures for adult stem cells, we have measured the post-rehydration membrane integrity (PRMI) of two passages, Passage-0 (P0) and Passage-1 (P1), of human adipose-derived stem cells (ASCs). ASCs were dried using a convective stage at three different drying rates (slow, moderate and rapid) in D-PBS with trehalose (50 mM) and glycerol (384 mM). ASCs were incubated in the drying media for 30 mins prior to drying at the prescribed rate on the convective stage for 30 mins. After drying, the ASCs were stored for 48 hrs in three different conditions: i) at ambient temperature, ii) in plastic bags at ambient temperature and iii) in vacuum sealed plastic bags at ambient temperature. PRMI was assessed after incubating the rehydrated ASCs with stromal medium for a further 48 hrs. Our measurements show that the PRMI of ASCs was: i) higher when ASCs were dried slowly; ii) increased when they were stored in vacuum as opposed to at ambient or in plastic bags; and iii) decreased with increasing passage of ASCs, i.e. under similar drying and storage conditions P0 ASCs had higher PRMI than P1 ASCs. Our results suggest that the best PRMI (37% for P0 ASCs and ~14% for P1 ASCs) can be achieved when the ASCs were dried slowly and stored in vacuum.

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Application of Nanomaterials in Regulating the Fate of Adipose-derived Stem Cells

Current Stem Cell Research & Therapy ◽

10.2174/1574888x15666200502000343 ◽

2021 ◽

Vol 16 (1) ◽

pp. 3-13

Author(s):

Lang Wang ◽

Yong Li ◽

Maorui Zhang ◽

Kui Huang ◽

Shuanglin Peng ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Stem Cell Research ◽

Adult Stem Cells ◽

Adipose Derived Stem Cells ◽

Proliferation And Differentiation ◽

Recent Developments ◽

Good Biocompatibility ◽

Regulatory Effects ◽

New Treatment

Adipose-derived stem cells are adult stem cells which are easy to obtain and multi-potent. Stem-cell therapy has become a promising new treatment for many diseases, and plays an increasingly important role in the field of tissue repair, regeneration and reconstruction. The physicochemical properties of the extracellular microenvironment contribute to the regulation of the fate of stem cells. Nanomaterials have stable particle size, large specific surface area and good biocompatibility, which has led them being recognized as having broad application prospects in the field of biomedicine. In this paper, we review recent developments of nanomaterials in adipose-derived stem cell research. Taken together, the current literature indicates that nanomaterials can regulate the proliferation and differentiation of adipose-derived stem cells. However, the properties and regulatory effects of nanomaterials can vary widely depending on their composition. This review aims to provide a comprehensive guide for future stem-cell research on the use of nanomaterials.

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Insufficient Stability of Clavulanic Acid in Widely Used Child-Appropriate Formulations

Antibiotics ◽

10.3390/antibiotics10020225 ◽

2021 ◽

Vol 10 (2) ◽

pp. 225

Author(s):

Ines Mack ◽

Mike Sharland ◽

Janneke M. Brussee ◽

Sophia Rehm ◽

Katharina Rentsch ◽

...

Keyword(s):

Ambient Temperature ◽

Clavulanic Acid ◽

Storage Conditions ◽

Essential Medicines ◽

Active Components ◽

Middle Income ◽

Ambient Temperatures ◽

Essential Medicines List ◽

Dispersible Tablets ◽

The Stability

Amoxicillin-clavulanic acid (AMC) belongs to the WHO Essential Medicines List for children, but for optimal antimicrobial effectiveness, reconstituted dry powder suspensions need to be stored in a refrigerated environment. Many patients in low- and middle-income countries who are sold AMC suspensions would be expected not to keep to the specified storage conditions. We aimed to assess the stability of both ingredients in liquid formulations and dispersible tablets, combined with nationally representative data on access to appropriate storage. Degradation of amoxicillin (AMX) and clavulanic-acid (CLA) was measured in suspensions and dispersible tablets commercially available in Switzerland at different ambient temperatures (8 °C vs. 28 °C over 7 days, and 23 °C vs. 28 °C over 24 h, respectively). Data on access to refrigeration and electricity were assessed from the USAID-funded Demographic and Health Survey program. In suspensions, CLA degraded to a maximum of 12.9% (95% CI −55.7%, +29.9%) at 8°C and 72.3% (95% CI −82.8%, −61.8%) at a 28 °C ambient temperature during an observation period of 7 days. Dispersible tablets were observed during 24 h and CLA degraded to 15.4% (95% CI −51.9%, +21.2%) at 23 °C and 21.7% (−28.2%, −15.1%) at a 28 °C ambient temperature. There is relevant degradation of CLA in suspensions during a 7-day course. To overcome the stability challenges for all active components, durable child-appropriate formulations are needed. Until then, prescribers of AMC suspensions or pharmacists who sell the drug need to create awareness for the importance of proper storage conditions regarding effectiveness of both antibiotics and this recommendation should be reflected in the WHO Essential Medicines List for children.

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Dual Inhibition of Activin/Nodal/TGF-βand BMP Signaling Pathways by SB431542 and Dorsomorphin Induces Neuronal Differentiation of Human Adipose Derived Stem Cells

Stem Cells International ◽

10.1155/2016/1035374 ◽

2016 ◽

Vol 2016 ◽

pp. 1-13 ◽

Cited By ~ 13

Author(s):

Vedavathi Madhu ◽

Abhijit S. Dighe ◽

Quanjun Cui ◽

D. Nicole Deal

Keyword(s):

Stem Cells ◽

Nervous System ◽

Neuronal Differentiation ◽

Adult Stem Cells ◽

Embryonic Stem ◽

Bmp Signaling ◽

Specific Marker ◽

Adipose Derived Stem Cells ◽

Neuronal Marker ◽

Dual Inhibition

Damage to the nervous system can cause devastating diseases or musculoskeletal dysfunctions and transplantation of progenitor stem cells can be an excellent treatment option in this regard. Preclinical studies demonstrate that untreated stem cells, unlike stem cells activated to differentiate into neuronal lineage, do not survive in the neuronal tissues. Conventional methods of inducing neuronal differentiation of stem cells are complex and expensive. We therefore sought to determine if a simple, one-step, and cost effective method, previously reported to induce neuronal differentiation of embryonic stem cells and induced-pluripotent stem cells, can be applied to adult stem cells. Indeed, dual inhibition of activin/nodal/TGF-βand BMP pathways using SB431542 and dorsomorphin, respectively, induced neuronal differentiation of human adipose derived stem cells (hADSCs) as evidenced by formation of neurite extensions, protein expression of neuron-specific gamma enolase, and mRNA expression of neuron-specific transcription factors Sox1 and Pax6 and matured neuronal marker NF200. This process correlated with enhanced phosphorylation of p38, Erk1/2, PI3K, and Akt1/3. Additionally,in vitrosubcutaneous implants of SB431542 and dorsomorphin treated hADSCs displayed significantly higher expression of active-axonal-growth-specific marker GAP43. Our data offers novel insights into cell-based therapies for the nervous system repair.

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Drying and storage of Eugenia involucrata DC. seeds

Scientia Agricola ◽

10.1590/s0103-90162003000300009 ◽

2003 ◽

Vol 60 (3) ◽

pp. 471-475 ◽

Cited By ~ 23

Author(s):

Angela Maria Maluf ◽

Denise Augusta Camargo Bilia ◽

Claudio José Barbedo

Keyword(s):

Moisture Content ◽

Water Content ◽

Cold Storage ◽

Native Species ◽

Plastic Bags ◽

Physiological Quality ◽

Drying Rates ◽

And Storage ◽

Water Contents

The physiological quality of seeds of native species is important to produce healthy saplings and therefore guarantee the success of programs to recover disturbed vegetation. This reinforces the necessity for investigating the physiological quality of those seeds. To evaluate the effects of different drying rates on the germination, moisture content and storability of Eugenia involucrata diaspores, mature fruits collected at Mogi Guaçu, SP, Brazil had their epi- and mesocarps removed by washing and were dried at 30, 40 or 50ºC until their water content was reduced from 57% (fresh diaspores) to 13% (final drying), totaling six drying levels. In a second experiment, diaspores had their moisture content reduced from 57% to 49%, at 30ºC, totaling six drying levels (0h, 1h, 2h, 3h, 4h and 5h), and were kept for 180 days in plastic bags under cold storage. The drying rate had no effect on tolerance to desiccation by E. involucrata diaspores; water contents lower than 51% decreased both germinability and storability. Diaspores can be stored for up to 180 days as long as their water content is reduced to 53% and they are kept inside plastic bags under cold storage.

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DEVELOPMENT OF DAMAGE IN THE BUDS DURING STORAGE OF IRRADIATED ONIONS

Canadian Journal of Plant Science ◽

10.4141/cjps72-133 ◽

1972 ◽

Vol 52 (5) ◽

pp. 817-826 ◽

Cited By ~ 3

Author(s):

NAOMI TEMKIN-GORODEISKI ◽

R. S. KAHAN ◽

R. PADOVA

Keyword(s):

Radiation Dose ◽

Ambient Temperature ◽

Shelf Life ◽

Storage Temperature ◽

Low Doses ◽

Ambient Temperatures ◽

Dormancy Period ◽

Growth Differences ◽

Effects Of Radiation ◽

And Storage

Darkening of onion buds due to irradiation was investigated during three seasons on three cultivars of onion, Riverside, Egyptian, and Grano. The effects of radiation dose (0.7–80 krads), delay in irradiation after harvest, and storage temperature were studied. No darkening occurred during storage of up to 8 months at 0 C, though slight darkening sometimes appeared during subsequent shelf life. At ambient temperatures (10–30 C) three types of radiation damage were found. The onset of darkening occurred not earlier than 2 months after irradiation. With all doses above 1.0 krad there was 100% incidence of darkening after about 5 months storage at ambient temperature. The intensity of darkening increased with length of storage. Very low doses (0.7–1.0 krad) did not prevent sprouting but caused slight darkening. Doses between 2 and 80 krads completely inhibited external sprouting; however, if delayed till 3 months postharvest, these doses failed to prevent sprouting, but did cause severe darkening, which was not dependent on the dose. There was no correlation between the delay in irradiation after harvest and the intensity or incidence of darkening. The length of the dormancy period varied in different years, apparently as a result of different conditions prevailing during crop growth. Differences in the intensity of darkening in different years seem to be connected with this phenomenon.

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Adipose-Derived Stem Cells in Tissue Regeneration: A Review

ISRN Stem Cells ◽

10.1155/2013/713959 ◽

2013 ◽

Vol 2013 ◽

pp. 1-35 ◽

Cited By ~ 71

Author(s):

Patricia Zuk

Keyword(s):

Stem Cells ◽

Los Angeles ◽

Tissue Regeneration ◽

Adult Stem Cells ◽

Adipose Derived Stem Cells ◽

Induced Pluripotent Cells ◽

Initial Description ◽

Induced Pluripotent

In 2001, researchers at the University of California, Los Angeles, described the isolation of a new population of adult stem cells from liposuctioned adipose tissue. These stem cells, now known as adipose-derived stem cells or ADSCs, have gone on to become one of the most popular adult stem cells populations in the fields of stem cell research and regenerative medicine. As of today, thousands of research and clinical articles have been published using ASCs, describing their possible pluripotency in vitro, their uses in regenerative animal models, and their application to the clinic. This paper outlines the progress made in the ASC field since their initial description in 2001, describing their mesodermal, ectodermal, and endodermal potentials both in vitro and in vivo, their use in mediating inflammation and vascularization during tissue regeneration, and their potential for reprogramming into induced pluripotent cells.

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401 MIGRATION AND THERAPEUTIC POTENTIAL OF PORCINE ADULT ADIPOSE-DERIVED MESENCHYMAL STEM CELLS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab401 ◽

2010 ◽

Vol 22 (1) ◽

pp. 357 ◽

Cited By ~ 2

Author(s):

S. M. Wilson ◽

E. Monaco ◽

M. S. Goldwasser ◽

S. G. Clark ◽

W. L. Hurley ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Adult Stem Cells ◽

Bone Defects ◽

Adipose Derived Stem Cells ◽

Subcutaneous Adipose ◽

Time Point

Bone marrow is one current source of adult stem cells for therapeutic purposes; however, the magnitude and accessibility of subcutaneous adipose tissue in humans make it an attractive alternative. Numerous in vitro studies have been conducted to determine how these cells act in vitro, but it is imperative to determine the vast abilities of these cells in vivo. The objective of this study was to evaluate in vivo migration and bone healing ability after transplanting adipose-derived stem cells (ADSC) in a swine model. Adipose-derived stem cells were isolated from subcutaneous adipose tissue of adult Yorkshire pigs and cultured in vitro. At 80 to 90% confluence/passage 3, the cells were trypsinized and labeled in suspension with carboxyfluorescein succinimidyl ester (CFDA-SE). This project included 20 pigs weighing between 63.5 and 81.7 kg. Bilateral mandibular osteoectomies with 10-mm defects were performed on each pig. Of the 20 pigs, half received a treatment of 2.5 million CFDA-SE labeled stem cells administered directly into each defect (DI), and the remaining half received a treatment of approximately 5 million CFDA-SE labeled stem cells through an ear vein injection via catheter (EVI). The time points were 1 h and 2 and 4 wk, with 2 pigs per time with the DI and EVI treatments. Pigs were slaughtered at each time, and spleen, liver, lung, kidney, ear vein, blood, and mandible tissues were collected. Blood samples were collected from the jugular vein with EDTA and processed via flow cytometry after collection. Tissues were fixed in 10% buffered formalin for histology. Fluorescent microscopy (CFDA-SE excitation/emission is 492/517 nm) has confirmed that transplanted ADSC do indeed migrate to a site of injury or trauma. Labeled cells were also present in blood collected from the 1-h time point group. Currently, we have not seen the presence of labeled ADSC in the other tissues (spleen, liver, lung, and kidney) after the 1-h time point. We did observe that ADSC administered by DI and EVI were able to significantly heal and regenerate bone defects within 4 wk post-surgery (P < 0.05, ANOVA with F-test), in contrast to bone defects in pigs that did not receive cell injections (control). Evidence of ADSC-related healing and bone regeneration was evident by gross visualization, dual-energy x-ray absorptiometry (DXA) and micro computer tomography (microCT) analysis. The clinical implications of these results are significant for treating many diseases in which inflammation or defects exist, such as cardiac disease, neurological disease, or traumatic injuries to both soft and hard tissue. If the adult stem cells can be harvested from fat, encouraged to produce bone or cartilage, and then reinserted into defects, treatment protocols for trauma victims could be developed that would reduce the need for alternate harvesting techniques for bone. This work was support by a grant from the Illinois Regenerative Medicine Institute (IDPH # 63080017).

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381 LABELING AND ANALYSIS OF SWINE ADIPOSE-DERIVED STEM CELLS WITH CARBOXYFLUORESCEIN DIACETATE AND QUANTUM DOTS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab381 ◽

2010 ◽

Vol 22 (1) ◽

pp. 347

Author(s):

N. Cieslak ◽

A. Massie ◽

S. M. Wilson ◽

E. Monaco ◽

M. B. Wheeler

Keyword(s):

Stem Cells ◽

Quantum Dots ◽

Regenerative Medicine ◽

Quantum Dot ◽

In Vitro Culture ◽

Adult Stem Cells ◽

Attractive Alternative ◽

Adipose Derived Stem Cells

The quantity, accessibility, and abundance of subcutaneous adipose tissue in humans make it an attractive alternative to bone marrow as a source of adult stem cells for therapeutic purposes. Adult adipose-derived mesenchymal stem cells can differentiate into a variety of lineages including adipose, bone, cartilage, and muscle. In addition, the use of adult stem cells for regenerative medicine rather than those from embryos avoids concerns with ethics, safety, and immunology. One important issue is the ability to track the transplanted stem cells during the regeneration process to evaluate the stem cell-mediated healing. The objective of this study was to compare the efficiency, longevity, and intensity of carboxyfluorescein diacetate, succinimidyl ester (CFDA SE) and quantum dot nanocrystal (Qtracker™, Invitrogen, Carlsbad, CA, USA) labeled adipose-derived stem cells (ADSC) over an in vitro culture period of 4 weeks. Adipose-derived stem cells (6 x 106) previously isolated and frozen at -196°C were thawed and cultured in 75-cm3 flasks with 14 mL of DMEM. Cells were grown to 80% confluence and trypsinized. After trypsinization, the cells were divided into 4 treatments (3 x 106 cells per treatment). The treatments were (1) unlabeled control, (2) labeled with 30 μM CFDA SE, (3) labeled with 15 nM Qtracker™, and (4) labeled with 15 nM Qtracker™, following the Invitrogen Qtracker™ protocol. Cells (1 x 106) were removed from each treatment every week for 4 weeks and fixed in formalin for later analysis. When all the samples were collected, they were analyzed using flow cytometry. Data were analyzed via chi-square test. The percentage of cells labeled with CFDA SE and Qtracker™ was 99.35 and 98.46%, respectively, immediately after labeling. By 1 wk, the percentage of cells labeled with CFDA SE and Qtracker™ had deceased (P < 0.01) to 0.11 and 1.48%, respectively. The CFDA SE-labeled cell percentages had decreased (P < 0.01) to 0% at 2, 3, and 4 wk, respectively. The Qtracker™-labeled cells also decreased (P < 0.01) to 0.745, 1.69 and 0.45% at 2, 3, and 4 wk, respectively. The high rate of cell division of these cells in vitro might be responsible for the rapid loss of both labels during the first week of culture. Previous results from our lab have shown that the CFDA SE is retained in the cells for up to 6 wk in vivo (Lima AS et al. 2006 Reprod. Fertil. Dev. 18, 208). Similar studies need to be done with the quantum dot-labeled cells to determine the Qtracker™ label’s longevity in vivo. In conclusion, quantum dots can be used to label ADSC, in vitro, for at least 4 wk, albeit at much lower levels than those observed during the week following labeling. Determination of a suitable label for high-percentage porcine ADSC labeling during long-term in vitro culture remains to be completed. This research was supported by the Intel Scholar’s Program and the Illinois Regenerative Medicine Institute.

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Effect of Various Freezing Parameters on the Immediate Post-Thaw Membrane Integrity of Adipose Tissue Derived Adult Stem Cells

Biotechnology Progress ◽

10.1021/bp050007q ◽

2005 ◽

Vol 21 (5) ◽

pp. 1511-1524 ◽

Cited By ~ 50

Author(s):

S. Thirumala ◽

S. Zvonic ◽

E. Floyd ◽

J.M. Gimble ◽

R.V. Devireddy

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Adult Stem Cells ◽

Membrane Integrity

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Impact of cultivation strategy, freeze-drying process, and storage conditions on survival, membrane integrity, and inactivation kinetics of Bifidobacterium longum

Folia Microbiologica ◽

10.1007/s12223-020-00815-3 ◽

2020 ◽

Vol 65 (6) ◽

pp. 1039-1050

Author(s):

Regina Haindl ◽

Alexandra Neumayr ◽

Anika Frey ◽

Ulrich Kulozik

Keyword(s):

Freeze Drying ◽

Bifidobacterium Longum ◽

Membrane Integrity ◽

Storage Conditions ◽

Inactivation Kinetics ◽

Whole Process ◽

Cultivation Strategy ◽

And Storage ◽

Kinetics Of ◽

Shelf Temperature

AbstractBifidobacterium longum, one of the main microorganisms in the human gut, is used as an adjunct to lactic acid starter cultures or sold as a probiotic product. Therefore, Bifidobacterium longum cell suspensions get freeze-dried with protective additives to prevent activity losses. To date, investigations covering growth and inactivation kinetics of Bifidobacterium longum during the whole process (cultivation, drying, and storage) have been lacking. In this study, the effect of cultivation conditions and shelf temperature as well as the influence of protectants (maltodextrin, glucitol, trehalose) at various concentrations on cell survival during freeze-drying was assessed. Drying was followed by a storage at + 4 °C and + 20 °C for 70 days to evaluate inactivation kinetics. The impact of the different factors was assessed by measuring surival rate and residual moisture content at various points of time over the whole process. In parallel cell membrane integrity and glass transition were determined to reveal inactivation effects. Cultivation strategy had a strong influence on survival with a huge potential for process improvement. A pH of 6.0 at the growth optimum of the strain provides better conditions regarding cell survival after drying than free acidification (non-regulated pH conditions). During the drying step, membrane leakage due to the removal of water is the main reason for the inactivation in this process step. In this study, the highest survival of 49% was obtained with cells dried at + 35 °C shelf temperature with an addition of maltodextrin (75% bacterial dry matter, w/w). The results show that Bifidobacterium longum cells are mostly inactivated during drying, whereas storage conditions at + 4 °C with an addition of 75% BDM maltodextrin relative to bacterial dry mass prevent cell loss completely.

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