Scellive tissue preservation solution

Manufactured by Singleron Biotechnologies

Sourced in China

SCelLiveTM Tissue Preservation Solution is a laboratory product designed to maintain the viability of tissue samples during storage and transportation. The solution is formulated to preserve the structural integrity and cellular function of tissue specimens, enabling researchers to conduct subsequent analyses and experiments.

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13 protocols using scellive tissue preservation solution

Single-cell RNA Sequencing of Spleen Tissue

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The fresh spleen tissues were stored in the sCelLive™ Tissue Preservation Solution (Singleron, China) on ice after the surgery within 30 min, and then digested with 3 mL sCelLive™ Tissue Dissociation Solution (Singleron) by PythoN™ Tissue Dissociation System (Singleron) at 37 °C for 15 min. The cell suspension was collected and filtered through a 40-micron sterile strainer. Afterwards, the red blood cell lysis buffer (RCLB, Singleron) was added and incubated at room temperature for 5-8 min. The mixture was then centrifuged to remove supernatant and suspended softly with PBS.
Single-cell suspensions (2 × 10⁵ cells/mL) with PBS were loaded onto a microwell chip using the Matrix® Single Cell Processing System (Singleron). Barcoding beads are subsequently collected from the microwell chip, followed by reverse transcription of the mRNA captured by the beads to obtain cDNA for PCR amplification. The amplified cDNA is then fragmented and ligated with sequencing adapters. The scRNA-seq libraries were constructed according to the protocol of the Single Cell RNA Library Kits (Singleron). Individual libraries were diluted to 4 nM, pooled, and sequenced on an Illumina Novaseq 6000 (Illumina, USA) instrument with a 150 bp paired-end format. Full identified sequencing data are available in the gene expression omnibus (GEO) under accession number GSE234756.

Xiong F., Zhang C., Shang B., Zheng M., Wang Q., Ding Y., Luo J, & Li X. (2023). An mRNA-based broad-spectrum vaccine candidate confers cross-protection against heterosubtypic influenza A viruses. Emerging Microbes & Infections, 12(2), 2256422.

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Single-cell isolation from cervical lesion

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After small cervical lesion tissues and blood samples were collected, the fresh tissue samples were immediately stored in the sCelLiVE^® Tissue Preservation Solution (Singleron, Nanjing, China) on ice. The tissues were cut into small tissue pieces and were transferred to a 15-ml centrifuge tube, followed by digestion using sCelLiVE^® Tissue Dissociation Solution (Singleron) at 37°C for 15 min with shaking. The samples were then filtered using 40-µm sterile strainers and centrifuged at 1,000 rpm at 4°C for 5 min. Next, 2 ml GEXSCOPE^® red blood cell lysis bufier (RCLB, Singleron) was added to lyse the red blood cells for 10 min. Finally, the single-cell suspension was collected after re-suspension with phosphate-buffered saline (PBS), and trypan blue (Sigma) staining was used to calculate cell activity and cell count under a microscope.

Liu X., Yu B., Gao F., Jing P., Zhang P., Zheng G, & Zhang X. (2024). Chemical immune conization of precancerous cervical lesions awakens immune cells and restores normal HPV negative and abnormal proliferation. Frontiers in Immunology, 14, 1259723.

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Single-Cell Isolation from Fibrotic and Healthy Livers

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The livers obtained from fibrotic liver of S. japonicum-induced mouse and healthy liver of normal mouse. The fresh tissues were stored in the sCelLive™ Tissue Preservation Solution (Singleron) on ice as quickly as possible. After washed with Hanks Balanced Salt Solution (HBSS) for three times, the specimens were minced into small pieces. To dissociated into single-cell suspensions, the specimens then digested with 3 mL sCelLive™ Tissue Dissociation Solution (Singleron) by Singleron PythoN™ Tissue Dissociation System at 37°C for 15 min. After filtered through a 40-micron sterile strainer, the cell was added the GEXSCOPE^® red blood cell lysis buffer (RCLB, Singleron), which was incubated at room temperature for 5-8 min to remove red blood cells. Centrifugation was performed at 300 × g 4°C for 5 mins to remove supernatant and single cells were resuspended softly in PBS. Finally, the cell viability was evaluated microscopically through staining with Trypan Blue.

Zhang Y., Li J., Li H., Jiang J., Guo C., Zhou C., Zhou Z, & Ming Y. (2022). Single-cell RNA sequencing to dissect the immunological network of liver fibrosis in Schistosoma japonicum-infected mice. Frontiers in Immunology, 13, 980872.

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Single-cell Isolation from Fresh Tissues

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The fresh tissues were preserved in the sCelLiVE Tissue Preservation Solution (Singleron) on ice within 30 min post-surgery. Following this, the specimens underwent a triple wash with Hanks Balanced Salt Solution, were minced into small pieces, and subsequently digested with 3 mL of sCelLiVE Tissue Dissociation Solution (Singleron) using the Singleron PythoN Tissue Dissociation System at 37°C for 15 min. The resulting cell suspension was collected and passed through a 40 micron sterile strainer. Next, GEXSCOPE red blood cell lysis buffer (RCLB, Singleron) was introduced to the cell suspension. This mixture, in a ratio of one part cell to two parts RCLB by volume, was incubated at room temperature for 5–8 min to eliminate red blood cells. After incubation, the mixture underwent centrifugation at 300×g at 4°C for 5 min to remove the supernatant. The remaining pellet was gently resuspended in PBS.

Long X., Zhang Z., Li Y., Deng K., Gao W., Huang M., Wang X., Lin X., She X., Zhao Y., Zhang M., Huang C., Wang S., Du Y., Du P., Chen S., Liu Q, & Wu M. (2024). ScRNA-seq reveals novel immune-suppressive T cells and investigates CMV-TCR-T cells cytotoxicity against GBM. Journal for Immunotherapy of Cancer, 12(4), e008967.

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Ascites Single Cell Processing

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After sample extraction (Figures 1C, D), the fresh cells samples were immediately stored in the sCelLiVE® Tissue Preservation Solution (Singleron) on ice. The sample of ascites were transferred to a 15-ml centrifuge tube. The samples were then filtered with 40 µm sterile strainers, and centrifuged at 1,000 rpm at 4°C for 5 min. Next, 2 ml GEXSCOPE® red blood cell lysis buffer (RCLB, Singleron) was added to lyse the red blood cells for 10 min. Finally, the single cell suspension was collected after re-suspension with PBS, and trypan blue (Sigma) staining was used to calculate cell activity and cell count under a microscope (7 (link)).

Yu B., Jing P., Gao F., Zhang P., Zheng G, & Zhang X. (2024). Effect of sodium stibogluconate in recruiting and awakening immune cells in the pleural fluid of pancreatic cancer: preparation for immunotherapy. Frontiers in Immunology, 14, 1315468.

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Tissue Dissociation and Single-Cell Isolation

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Fresh tissues were stored on ice in the sCelLive™ Tissue Preservation Solution (Singleron) within 30 min after surgery. Hanks Balanced Salt Solution (HBSS) was used three times to wash the specimens, mince them into small pieces, and digest them in 3 mL sCelLive™ Tissue Dissociation Solution (Singleron) by Singleron PythoN™ Tissue Dissociation System at 37°C for 15 min. A 40-micron sterile strainer was used to collect and filter the cell suspension. After adding the GEXSCOPE^® red blood cell lysis buffer (RCLB, Singleron), the mixture [Cell: RCLB = 1:2 (volume ratio)] was incubated for 5–8 min at room temperature to remove red blood cells. After centrifuging at 300 × g 4°C for 5 min, the mixture was suspended in PBS softly after centrifugation. Finally, Trypan Blue staining was applied to the samples, and cell viability was assessed microscopically.

Huang X., Lu Z., Jiang X., Zhang Z., Yan K, & Yu G. (2023). Single-cell RNA sequencing reveals distinct tumor microenvironment of ground glass nodules and solid nodules in lung adenocarcinoma. Frontiers in Cell and Developmental Biology, 11, 1198338.

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Isolation of Renal Cell Populations

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Within 30 min of procurement from both LN and normal donors, the renal tissue samples were preserved in the sCelLiveTM Tissue Preservation Solution (Singleron, China) on ice. The samples were then rinsed three times with Hanks Balanced Salt Solution, cut into small pieces, and digested using 3 mL of sCelLiveTM Tissue Dissociation Solution with the Singleron PythoN Tissue Dissociation System at 37°C for 15 min. The cell mixture was gathered and passed through a 40‐micron sterile filter to separate the cells. After that, the GEXSCOPE red blood cell lysis buffer (RCLB, Singleron) was added to the mixture in a 1:2 (volume) ratio with the cells and incubated at room temperature for 5–8 min to eliminate red blood cells. The mixture was then centrifuged at 300xg and 4°C for 5 min to remove the supernatant and resuspended softly in phosphate buffered saline (PBS) (HyClone, USA).

Tang Y., Zhang Y., Li X., Xu R., Ji Y., Liu J., Liu J., Zhuang Q, & Zhang H. (2023). Immune landscape and the key role of APOE+ monocytes of lupus nephritis under the single‐cell and spatial transcriptional vista. Clinical and Translational Medicine, 13(4), e1237.

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Tissue Dissociation and Cell Viability

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Fresh tissues were stored in sCelLiveTM Tissue Preservation Solution (Singleron, China) on ice immediately after resection. The tissues were washed with HBSS three times and then digested with 2 mL sCelLiveTM Tissue Dissociation Solution (Singleron) at 37 °C for 15 min. To remove red blood cells, red blood cell lysis buffer (2 mL) was added to the cells and incubated at 25 °C for 10 min. The solution was then centrifuged at 500 × g for 5 min and suspended in PBS. Trypan blue (Sigma) staining was used to evaluate cellular viability under a microscope.

Zou Y., Ye F., Kong Y., Hu X., Deng X., Xie J., Song C., Ou X., Wu S., Wu L., Xie Y., Tian W., Tang Y., Wong C., Chen Z., Xie X, & Tang H. (2022). The Single‐Cell Landscape of Intratumoral Heterogeneity and The Immunosuppressive Microenvironment in Liver and Brain Metastases of Breast Cancer. Advanced Science, 10(5), 2203699.

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Single-cell RNA-seq of Mouse Kidneys

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The kidney samples of mice in different feeding groups were collected for the scRNA-seq assay. Briefly, the kidney tissues were preserved in sCelLiveTM tissue preservation solution (Singleron Biotechnologies, Nanjing, China). Then, the tissues were separated into single-cell suspensions using Singleron Python Python™ Automated Tissue Separator (Singleron Biotechnologies) and sCelLive™ Tissue Isolation Mix (Singleron Biotechnologies) following the manufacturer’s instructions. Finally, cell viability was assessed under a microscope after staining with Trypan Blue (Sigma).

Xu W., Ye S., Liu W., Guo H., Zhang L., Wei S., Anwaier A., Chang K., Malafaia G., Zhang H., Ye D, & Wei G. (2024). Single-cell RNA-seq analysis decodes the kidney microenvironment induced by polystyrene microplastics in mice receiving a high-fat diet. Journal of Nanobiotechnology, 22, 13.

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Isolation of Testicular Cell Samples

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Three unilaterally decapsulated testes were dissected from 3 adult WT and 3 adult KO mice. The fresh testicular tissues were stored in the sCelLiveTM Tissue Preservation Solution (Singleron Biotechnologies) on ice within 30 min after the surgery. The tissues were washed with Hanks’ Balanced Salt Solution thrice and then digested with 2 ml sCelLiveTM Tissue Dissociation Solution (Singleron Biotechnologies) using the Singleron PythoN™ Automated Tissue Dissociation System (Singleron Biotechnologies) at 37°C for 15 min. The tissue lysate was centrifuged at 500 × g for 5 min, and the pellet was suspended gently with PBS. The samples were stained with trypan blue (T8154; Sigma), and the cellular viability was evaluated microscopically. Apoptotic or dead cells were discarded.

Hong S., Shen X., Luo C, & Sun F. (2022). Comparative analysis of the testes from wild-type and Alkbh5-knockout mice using single-cell RNA sequencing. G3: Genes|Genomes|Genetics, 12(8), jkac130.

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