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Table 1 Summary of single-cell isolation protocols for nucleus pulposus in reported studies using scRNA-seq technique

From: Systematic study of single-cell isolation from musculoskeletal tissues for single-sell sequencing

Reference

Classification of IVD

Species

Treatment

Dead cell removal

Profiling cellular types

Gan Y, et al. [10]

Pfirrmann grade I

Human

TrypLE Express for 30 min, 0.2% pronase (Sigma-Aldrich, USA) for 60 min, and 0.012% collagenase II (Sigma-Aldrich, USA) for 2 ~ 4 h, sequentially

Yes

Chondrocyte, Notochord, Nucleus pulposus progenitor cells, Stromal cell, Pericyte, Endothelial cell and blood cell

Fernandes LM, et al. [11]

Thompson grade 1 or 2

Human

Pronase for 1 h and then collagenase P overnight

No

Culture cells for scRNA-seq

Cultured nucleus pulposus cells

Calió M, et al. [12]

 

Bovine

0.19% Pronase for 1 h and then 260 IU/mg Collagenase II overnight

No

Nucleus pulposus cells, uncharacterized cell subset, monocyte derived dendritic cells

Zhang Y, et al. [13]

Pfirrmann grade I-IV

Human

GEXSCOPETM Tissue Dissociation Solution for 15 min. (Included in the GEXSCOPE® Single Cell RNA library Kit)

No

Chondrocytes, macrophages and endothelial cells

Ji Tu, et al. [14]

Pfirrmann grade II-V

Human

500 U/mL collagenase I, 150 U/mL collagenase II, 50 U/mL collagenase IV, 0.1 mg/ml hyaluronidase and 30 U/mL DNaseI for 95 min

Yes

Neucleus pulposus cells, natural killer cells, monocytes, and T cells