702.B.3 URMC HTC Lung Tissue Digestion SOP + Worksheet 052820 v1

Lung MAP HTC - BioRepository for Investigation of Neonatal Diseases of the Lung (BRINDL) 702.B.3 Combined SOP and Worksheet HTC_Lung_Tissue_Digestion_0100616rev061220 Purpose and Scope of the Procedure or Laboratory Assay 1.To dissociate and isolate single cell suspensions from human lung samples to meet the standards and needs of the Human Tissue Core. 2.This protocol covers manual gross dissociation of pieces of lung tissue (lobes or cubes) followed by enzymatic digestion and freezer storage as mixed populations of filtered and counted cells for which viability has been determined. 3.A separate protocol details the technique of freezing, thawing and further identifying and sorting these mixed cell populations. Scientific Principles or Validation of Procedure 1.Fujino et al. Am J Respir Cell Mol Biol. 2012 Apr;46(4):422-30 2.Barkauskas, C. E., Cronce, M. J., Rackley, C. R., Bowie, E. J., Keene, D. R., Stripp, B. R., ... & Hogan, B. L. (2013). Type 2 alveolar cells are stem cells in adult lung. The Journal of clinical investigation, 123(7), 3025.