Scienceware flowmi 40 m cell strainers

Samples of the eutopic endometrium and ectopic lesions from patient with AM and eutopic endometrium from patient with hysteromyoma were isolated on the same day and transported rapidly to the research facility. Then, each sample was chopped into cubes of no more than 1 mm under the condition of 4℃, followed by digestion with collagenase, shaking evenly every 5 min. Next, the supernatant was removed after centrifugation at 300 relative centrifugal force (RCF) for 30 s at room temperature (RT), without disturbing the cell particles. BSA (400 µg/ml) prepared with 1 × PBS free with calcium and magnesium was added, after which the samples were centrifuged at 300 RCF for 5 min at RT, and the cell pellets were resuspended in 1 ml red blood cell lysate buffer, followed by incubation at 4℃ for 10 min. After red blood cell lysis, the samples were resuspended in 1 ml of BSA. Finally, the processed samples were filtered through Scienceware Flowmi 40-µm cell strainers (VWR). A haemocytometer and trypan blue staining were used to evaluate cell concentration and viability.

The single-cell isolation and sequencing were performed in Shanghai OE Biotech. Co. Ltd. (Shanghai, China). Samples from resection surgery were isolated and transported rapidly to the research facility. Each sample was minced on ice to less than 1 mm cubic pieces, and digested using collagenase type I (Gibco, Grand Island, NY, USA). Then, samples were centrifuged at 300 rcf for 30 s at room temperature and removed the supernatant without disturbing the cell pellet. Samples were washed with phosphate-buffered saline (PBS) containing 0.04% BSA (400 µg/mL) and then centrifugation was performed at 300 rcf for 5 min. The cell pellet was resuspended in red blood cell lysis buffer and incubated for 10 min at 4 °C and then resuspended in 1 mL PBS containing 0.04% BSA. Next, samples were filtered over Scienceware Flowmi 40 µm cell strainers (VWR). After tissue dissociation, cell concentration and cell viability were determined using the Trypan Blue staining method to assess the quality of the samples.