Characterization and modulation of human mesenchymal stem cell stress pathway response following hypothermic storage

Abstract

Human mesenchymal stem cell (hMSC) research has grown exponentially in the last decade. The ability to process and preserve these cells is vital to their use in stem cell therapy. As such, understanding the complex, molecular-based stress responses associated with biopreservation is necessary to improve outcomes and maintain the unique stem cell properties specific to hMSC. In this study hMSC were exposed to cold storage (4°C) for varying intervals in three different media. The addition of resveratrol or salubrinal was studied to determine if either could improve cell tolerance to cold. A rapid elevation in apoptosis at 1 hour post-storage as well as increased levels of necrosis through the 24 hours of recovery was noted in samples. The addition of resveratrol resulted in significant improvements to hMSC survival while the addition of salubrinal revealed a differential response based on the media utilized. Decreases in both apoptosis and necrosis together with decreased cell stress/death signaling protein levels were observed following modulation. Further, ER stress and subsequent Unfolded Protein Response (UPR) stress pathway activation was implicated in response to hMSC hypothermic storage. This study is an important first step in understanding hMSC stress responses to cold exposure and demonstrates the impact of targeted molecular modulation of specific stress pathways on cold tolerance thereby yielding improved outcomes. Continued research is necessary to further elucidate the molecular mechanisms involved in hypothermic-induced hMSC cell death. This study has demonstrated the potential for improving hMSC processing and storage through targeting select cell stress pathways.

Influence of Factors of Cryopreservation and Hypothermic Storage on Survival and Functional Parameters of Multipotent Stromal Cells of Placental Origin

“…Analysis of cellular characteristics after hypothermic storage at +4°C in the culture medium without additional components for 24 hours revealed significant decrease of the cell number ( Table 1 ), which corresponds with the literature [ 28 , 29 ]. Interestingly, all cells were distinctly located without forming conglomerates ( Fig 3c ) and adhered instead with poor distribution on the dish, slow proliferation and late monolayer formation on day 8 ( Fig 3d ).…”

“…As to hypothermic storage, different levels of success can be found in the literature, strongly depended on the cell type and composition of preservation media [ 19 , 32 , 33 ]. In summary, hypothermic storage of cells in suspension causes swelling of cells due to cold inactivation of membrane pumps, as well as activation of cellular stress pathways [ 28 , 34 ] resulting in rapid depletion of cellular energy reserves and loss of stored material. For our studied cell type, hypothermic storage in culture medium without the addition of colloids, antioxidants, and sugars does not allow necessary preservation outcome during this period, which is most likely connected to damages caused by hypothermia.…”

Clinically Relevant Solution for the Hypothermic Storage and Transportation of Human Multipotent Mesenchymal Stromal Cells

“…This highly effective complex solution, available on the market under the trademarks VIASPAN®, SPS-1® (Organ Recovery Systems, USA), and Belzer UW® Cold Storage Solution (Preservation Solutions Inc., USA), contains a large panel of components, which are aimed at providing an improved viability of organs prior to transplantation [12]. Several reports show its applicability for the hypothermic preservation of MSCs [17, 18]. However, the complex composition of this solution, which needs to be validated, according to clinical GMP standards for injectable devices, complicates its use as a vehicle solution for cell delivery proposes.…”

Alginate-Encapsulation for the Improved Hypothermic Preservation of Human Adipose-Derived Stem Cells

“…The examination of hypothermic preservation of MSCs has previously been reported on cells derived from different tissues and at different storage temperatures, including (a) bone marrow at 4°C [31][32][33][34][35][36][37][38], 24°C [33,35], ambient temperature [22,36], and 37°C [35]; (b) adipose tissue at 4°C [39,40], 8°C, 25°C, and 37°C [40]; and (c) umbilical cord at 4°C [41]. Although these studies presented promising findings, most had been conducted at 4°C.…”