Recent Advances and Future Direction in Lyophilisation and Desiccation of Mesenchymal Stem Cells

Bissoyi, Akalabya; Kumar, Awanish; Rizvanov, Albert A.; Несмелов, А. А.; Gusev, Oleg; Patra, Pradeep Kumar; Bit, Arindam

doi:10.1155/2016/3604203

Cited by 27 publications

(23 citation statements)

References 43 publications

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“…While cryopreservation allows for the long-term storage of tissue containing viable cells, the need for specialized equipment to maintain ultralow temperatures for shipment and storage limits the use of cryopreserved product to medical facilities that have deep freezers. 21 Advances in preservation techniques, however, have led to the development of a lyopreservation method that allows for viable tissues to be stored at room temperature. Using this lyopreservation technique, a new formulation of placental membrane, has recently been developed.…”

Section: Introductionmentioning

confidence: 99%

Wound Closure Outcomes Suggest Clinical Equivalency Between Lyopreserved and Cryopreserved Placental Membranes Containing Viable Cells

Ananian¹,

Davis

Johnson

et al. 2019

Advances in Wound Care

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Objective: To evaluate the clinical outcomes of lyopreserved placental membrane containing viable cells (vLPM) in the treatment of nonhealing wounds of various etiologies, and to compare them to those previously reported for cryopreserved placental membrane containing viable cells (vCPM). Approach: Patients with nonhealing wounds who qualified to receive advanced wound therapies were consecutively enrolled and treated weekly with vLPM plus standard of care (SOC) at five centers. Data were de-identified and retrospectively analyzed. Outcomes included closure, time to closure, number of vLPM applications, and adverse events (AEs). Results: Seventy-eight patients with 98 wounds (41 diabetic foot ulcers [DFUs], 19 venous leg ulcers [VLUs], 10 surgical, and 28 others) with an average size of 13.3 cm 2 and 8.7 months duration were treated. Fifty-eight of the 98 wounds (59.2%) achieved complete closure with median time to closure of 63 days and 6 vLPM applications. The closure by wound etiology was 63% for DFUs, 47% for VLUs, 70% for surgical wounds, and 57% for other types of wounds. Similar closure rates have been previously demonstrated for vCPM. Wound duration was the main predictor of closure: 65.8% versus 30.0% (p = 0.004) closure was achieved for wounds of £12 and >12 months duration, respectively. There were no AEs related to vLPM application. Innovation: This is the first multicenter case series evaluating the clinical outcomes of vLPM in a real-world setting. Conclusion: These results support clinical equivalency between the two placental membrane formulations with the added convenience of room-temperature storage for vLPM, allowing it to be used in any wound-care setting.

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Section: Introductionmentioning

confidence: 99%

Wound Closure Outcomes Suggest Clinical Equivalency Between Lyopreserved and Cryopreserved Placental Membranes Containing Viable Cells

Ananian¹,

Davis

Johnson

et al. 2019

Advances in Wound Care

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“…As a non-reducing disaccharide, trehalose has a remarkably high glass transition temperatures (Tg) (Levine and Slade, 1988;Eroglu et al, 2009;Eroglu, 2010), which would support the resulting vitrified state at high temperatures. Therefore, trehalose has been extensively used for preservation of biological materials (Bissoyi et al, 2016). Experimental evidence suggest that trehalose exerts its protective effects (i) by stabilizing lipid membranes as a result of direct FIGURE 5 | Effect of sodium hydrogen sulfide (NaHS) and hydrogen sulfide (H 2 S) donor GYY4137 on ischemic human myoblasts.…”

Section: Discussionmentioning

confidence: 99%

Freeze-Dried Extracellular Vesicles From Adipose-Derived Stem Cells Prevent Hypoxia-Induced Muscle Cell Injury

Baradie

Nouh

O'Brien

et al. 2020

Front. Cell Dev. Biol.

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Cellular therapies have tremendous potential for the successful treatment of major extremity wounds in the combat setting, however, the challenges associated with transplanting stem cells in the prolonged field care (PFC) environment are a critical barrier to progress in treating such injuries. These challenges include not only production and storage but also transport and handling issues. Our goal is to develop a new strategy utilizing extracellular vesicles (EVs) secreted by stem cells that can resolve many of these issues and prevent ischemic tissue injury. While EVs can be preserved by freezing or lyophilization, both processes result in decrease in their bioactivity. Here, we describe optimized procedures for EVs production, isolation, and lyophilization from primary human adipose-derived stem cells (hADSCs). We compared two isolation approaches that were ultrafiltration (UF) using a tangential fluid filtration (TFF) system and differential ultracentrifugation (UC). We also optimized EVs lyophilization in conjunction with trehalose and polyvinylpyrrolidone 40 (PVP40) as lyoprotectants. Bioactivity of EVs was assessed based on reversal of hypoxia-induced muscle cell injury. To this end, primary human myoblasts were subjected to hypoxic conditions for 6 h, and then treated with hADSC-derived EVs at a concentration of 50 µg/mL. Subsequently, muscle cell viability and toxicity were evaluated using MTS and LDH assays, respectively. Overall, nanoparticle tracking data indicated that UF/TFF yields threefold more particles than UC. Lyophilization of EVs resulted in a significantly reduced number of particles, which could be attenuated by adding lyoprotections to the freeze-drying solution. Furthermore, EVs isolated by UF/TFF and freeze-dried in the presence of trehalose significantly increased viability (P < 0.0193). Taken together, our findings suggest that the isolation and preservation methods presented in this study may enhance therapeutic applications of EVs.

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“…Control cells can be used as positive and procedural controls for flow cytometric applications such as phenotyping of leukemia and lymphoma, immune monitoring and drug discovery. Scientists have been exploring cell preservation techniques to achieve both consistency and performance for a few decades (68,76,77,83). This workshop covered all current products as well as laboratory-developed approaches.…”

Section: Discussionmentioning

confidence: 99%

Cyt‐Geist: Current and Future Challenges in Cytometry: Reports of the CYTO 2018 Conference Workshops

Czechowska¹,

Lannigan

Wang

et al. 2019

Cytometry Pt A

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Recent Advances and Future Direction in Lyophilisation and Desiccation of Mesenchymal Stem Cells

Cited by 27 publications

References 43 publications

Wound Closure Outcomes Suggest Clinical Equivalency Between Lyopreserved and Cryopreserved Placental Membranes Containing Viable Cells

Wound Closure Outcomes Suggest Clinical Equivalency Between Lyopreserved and Cryopreserved Placental Membranes Containing Viable Cells

Freeze-Dried Extracellular Vesicles From Adipose-Derived Stem Cells Prevent Hypoxia-Induced Muscle Cell Injury

Cyt‐Geist: Current and Future Challenges in Cytometry: Reports of the CYTO 2018 Conference Workshops

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