Effects of carrier solutions on the viability and efficacy of canine adipose-derived mesenchymal stem cells

Abstract

Background Mesenchymal stem cells (MSCs) have favorable characteristics that render them a potent therapeutic tool. We tested the characteristics of MSCs after temporal storage in various carrier solutions, such as 0.9% saline (saline), 5% dextrose solution (DS), heparin in saline, and Hartmann’s solution, all of which are approved by the U.S. Food and Drug Administration (FDA). Phosphate-buffered saline, which does not have FDA approval, was also used as a carrier solution. We aimed to examine the effects of these solutions on the viability and characteristics of MSCs to evaluate their suitability and efficacy for the storage of canine adipose-derived MSCs (cADMSCs). Results We stored the cADMSCs in the test carrier solutions in a time-dependent manner (1, 6, and 12 h) at 4 °C, and analyzed cell confluency, viability, proliferation, self-renewability, and chondrogenic differentiation. Cell confluency was significantly higher in 5% DS and lower in phosphate-buffered saline at 12 h compared to other solutions. cADMSCs stored in saline for 12 h showed the highest viability rate. However, at 12 h, the proliferation rate of cADMSCs was significantly higher after storage in 5% DS and significantly lower after storage in saline, compared to the other solutions. cADMSCs stored in heparin in saline showed superior chondrogenic capacities at 12 h compared to other carrier solutions. The expression levels of the stemness markers, Nanog and Sox2, as well as those of the MSC surface markers, CD90 and CD105, were also affected over time. Conclusion Our results suggest that MSCs should be stored in saline, 5% DS, heparin in saline, or Hartmann’s solution at 4 °C, all of which have FDA approval (preferable storage conditions: less than 6 h and no longer than 12 h), rather than storing them in phosphate-buffered saline to ensure high viability and efficacy.

Refrigerated human mesenchymal stromal cells as an alternative to cryostorage for use in clinical investigation

“…In most of these studies, DMEM growth media was compared to crystalloid solutions ranging from 0.9% saline to PBS to Ringer's lactate and Plasma-Lyte. [26][27][28][29][30][31][32] Specialized storage media examined included HypoThermosol and buffered trehalose solutions. 33,34 With or without the addition of human serum albumin, the maximum duration studied was 4 days.…”

“…Previous studies investigated 4°C storage with a focus on transport. In most of these studies, DMEM growth media was compared to crystalloid solutions ranging from 0.9% saline to PBS to Ringer's lactate and Plasma‐Lyte 26–32 . Specialized storage media examined included HypoThermosol and buffered trehalose solutions 33,34 .…”

Optimal Intravenous Administration Procedure for Efficient Delivery of Canine Adipose-Derived Mesenchymal Stem Cells

“…We hypothesized three possible causes of poor cell-delivery efficiency during intravenous infusion: (1) acute cell death due to nutrient deprivation and hypoxic stress in the suspension solution or biomechanical stress during infusion; (2) cell adhesion to the walls of the apparatus, such as the infusion bag, syringe, or infusion tube; and (3) cell aggregation during storage and infusion. Temperature, solution type, and cell density of the cell preparation, as well as storage, can affect the metabolic activity of MSC, thus altering their viability and function [ 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 ]. It has been reported that the viability of hADSC in NS is better maintained at lower temperatures (4 °C) than at room temperature (37 °C) [ 42 , 43 ].…”

“…Suspension in DEX appeared to cause cell bursting and an increase in dead cells. Pal et al reported that DEX was suitable for preserving hBMSC [ 45 ], but similar studies with hUCMSC [ 46 ], hADSC [ 47 ], and cADSC [ 48 ] reported low viability with this solution. The mechanism responsible for the decreased viability is unknown, but may be related to factors other than pH or osmotic pressure [ 46 ].…”

Manufacturing Mesenchymal Stromal Cells for the Treatment of Osteoarthritis in Canine Patients: Challenges and Recommendations

“…Saline solution and 5%DS were able to maintain optimal viability and colony forming units ability up to 12hrs. Gene expression levels of CD105 and CD90 reached significantly higher expression in cells stored in PBS, while collagen type II alpha 1 chain (COL2A) and SRY-Box Transcription Factor 9 (Sox 9) were highly expressed in chondrogenically-differentiated MSCs that had been stored respectively in saline and HS (209). Based on this, it is important to carefully consider and test various cell carriers by testing their efficacy and stability on the final product of the manufacturing process.…”