HCA Data Explorer

Cryopreservation and post-thaw characterization of dissociated human islet cells

Updated November 10, 2023

This study aims to optimize the cryopreservation of dissociated islet cells and obtain functional cells that can be used in single-cell transcriptome studies on the pathology and treatment of diabetes. Using an iterative graded freezing approach we obtained viable cells after cooling in 10% dimethyl sulfoxide and 6% hydroxyethyl starch at 1 °C/min to –40 °C, storage in liquid nitrogen, rapid thaw, and removal of cryoprotectants by serial dilution. The expression of epithelial cell adhesion molecule declined immediately after thaw, but recovered after overnight incubation, while that of an endocrine cell marker (HPi2) remained high after cryopreservation. Patch-clamp electrophysiology revealed differences in channel activities and exocytosis of various islet cell types; however, exocytotic responses, and the biophysical properties of voltage-gated Na+ and Ca2+ channels, are sustained after cryopreservation. Single-cell RNA sequencing indicates that overall transcriptome and crucial exocytosis genes are comparable between fresh and cryopreserved dispersed human islet cells. Thus, we report an optimized procedure for cryopreserving dispersed islet cells that maintained their membrane integrity, along with their molecular and functional phenotypes. Our findings will not only provide a ready source of cells for investigating cellular mechanisms in diabetes but also for bio-engineering pseudo-islets and islet sheets for modeling studies and potential transplant applications.

Janet AW ElliottUniversity of Albertajanet.elliott@ualberta.ca
Yan Hang (Experimental Scientist)1
Xiao-Qing Dai (Experimental Scientist)2
Leah A Marquez-Curtis (Experimental Scientist)2
Janet AW Elliott (Principal Investigator)2
1Stanford University
2University of Alberta
Rachel Schwartz

To reference this project, please use the following link:

https://explore.data.humancellatlas.org/projects/8559a8ed-5d8c-4fb6-bde8-ab639cebf03c
None
None

For information regarding data sharing and data use, please see our Data Access Policy.

Analysis Portals

None

Project Label

CryoPancreaticIsletCellPatchSeq

Species

Homo sapiens

Sample Type

specimens

Anatomical Entity

pancreas

Organ Part

islet of Langerhans

Selected Cell Types

epithelial cell of pancreas

Disease Status (Specimen)

normal

Disease Status (Donor)

normal

Development Stage

human adult stage

Library Construction Method

Smart-seq2

Nucleic Acid Source

single cell

Paired End

true

Analysis Protocol

MultiSampleSmartSeq2_v2.2.6, SmartSeq2SingleSample_v5.1.5, analysis_protocol_1

File Format

6 file formats

Cell Count Estimate

189

Donor Count

3
asc189 file(s)bai189 file(s)bam189 file(s)fastq.gz378 file(s)loom1 file(s)xlsx2 file(s)