Background: Formalin has been widely used by pathology laboratories. Its carcinogenicity has led researchers to explore formalin substitutes. Streck Cell Preservative (SCP) is a formalin-free preservative that can preserve cellular antigens. This study was undertaken to investigate the effects of cell preservation using SCP on nucleic acid amplification, hybridization, and next-generation sequencing (NGS) as compared to control frozen cells and cells fixed in the traditional cell and tissue fixative, 10 % neutral buffered formalin (NBF).
Findings: The breast cancer cell line, SKBR-3, was used as a model system. Prior to nucleic acid extraction and fluorescence in situ hybridization (FISH), cells were fixed in SCP or NBF overnight at room temperature with frozen cells in parallel. Analysis showed that similar DNA extraction yields and amplification profiles determined by PCR in SCP preserved cells and control frozen cells, whereas NBF preserved cells had decreased DNA yield and impaired PCR amplification. Molecular cytogenetic studies by FISH technique indicated that the ratios of ERBB2 (HER-2/neu) signals to the chromosome 17 centromere (CEP17) were comparable for frozen cells and SCP preserved cells. The fluorescence images of both SCP fixed and control frozen cells were also clear and comparable. On the contrary, the same analysis was unsuccessful with NBF preserved cells due to poor hybridization quality. Our data also demonstrated that SCP had negligible effect on NGS testing.
Conclusion: We conclude that SCP can be used as an alternative to NBF as a preservative for maintaining the integrity of nucleic acids for nucleic acid amplification, sequencing and FISH analysis.
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http://dx.doi.org/10.1186/s13104-015-1725-4 | DOI Listing |
Surgery
January 2025
Department of Surgery, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH.
Introduction: In response to blood shortages, providers face pressure to conserve blood. No metrics exist to calculate transfusion utility. We describe characteristics of survivors after high-volume resuscitation and evaluate transfusion utility in low-volume and high-volume resuscitation.
View Article and Find Full Text PDFCryobiology
January 2025
Animal Physiology Division, ICAR-National Institute of Animal Nutrition and Physiology, Adugodi, Bangalore, India-560030.
The application of spermatogonial stem cells (SSC) will be more effective and feasible following the successful cryopreservation and transfer of SSCs in livestock. Like other cells, SSCs are also sensitive to cryoinjury; hence composition of the cryomedia and freezing protocols need to be optimized. The present study aims to optimising the best freezing rates by minimising the ice crystallization and dehydration effect in order to maximize the post-thaw SSCs survivability and stemness characteristics.
View Article and Find Full Text PDFInt J Mol Sci
January 2025
Advanced Biological Therapy Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, Spain.
Platelet-Rich Plasma (PRP) is a biological treatment widely used in regenerative medicine for its restorative capacity. Although PRP is typically applied at the time of obtention, long-term storage and preservation could enhance its versatility and clinical applications. The objective of this study was to evaluate the effect of long-term freezing on PRP.
View Article and Find Full Text PDFGenes (Basel)
January 2025
Department of Biological Sciences, Marquette University, Milwaukee, WI 53233, USA.
Background/objectives: Cold stress poses a significant threat to Asian rice cultivation, disrupting important physiological processes crucial for seedling establishment and overall plant growth. It is, thus, crucial to elucidate genetic pathways involved in cold stress tolerance response mechanisms.
Methods: We mapped , a ()-type homolog of rice, to a low-temperature seedling survivability (LTSS) QTL and used genomics, molecular genetics, and physiological assays to assess its role in plant resilience against low-temperature stress.
Animals (Basel)
January 2025
Veterinary Clinic for Reproductive Medicine and Neonatology, Justus-Liebig-University of Giessen, 35392 Giessen, Germany.
Cryopreservation can adversely affect sperm motility, structural integrity, and fertilization ability. This study investigated the effects of MitoQ and antifreeze protein III (AFP III) on frozen-thawed semen from eight adult dogs using a Tris-fructose extender. Ejaculates were divided and diluted with a standard Tris-fructose-egg yolk extender containing MitoQ (200 nM/mL) and AFP III (0.
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