Optimizing the transport and storage conditions of current Good Manufacturing Practice -grade human umbilical cord mesenchymal stromal cells for transplantation (HUC-HEART Trial)

Ferda Topal Celikkan; Ceren Mungan; Merve Sucu; A Tulga Ulus; Ozgur Cinar; Ezgi Gokpinar Ili; Alp Can

doi:10.1016/j.jcyt.2018.10.010

Optimizing the transport and storage conditions of current Good Manufacturing Practice -grade human umbilical cord mesenchymal stromal cells for transplantation (HUC-HEART Trial)

Cytotherapy. 2019 Jan;21(1):64-75. doi: 10.1016/j.jcyt.2018.10.010. Epub 2018 Nov 16.

Authors

Ferda Topal Celikkan¹, Ceren Mungan², Merve Sucu², A Tulga Ulus³, Ozgur Cinar¹, Ezgi Gokpinar Ili⁴, Alp Can⁵

Affiliations

¹ Department of Histology and Embryology, Faculty of Medicine, Ankara University, Ankara, Turkey.
² Ankara University Biotechnology Institute, Ankara, Turkey.
³ Division of Cardiovascular Surgery, Acibadem Hospital, Ankara, Turkey.
⁴ Department of Medical Genetics, Faculty of Medicine, Ankara University, Ankara, Turkey.
⁵ Department of Histology and Embryology, Faculty of Medicine, Ankara University, Ankara, Turkey. Electronic address: alpcan@medicine.ankara.edu.tr.

PMID: 30455106
DOI: 10.1016/j.jcyt.2018.10.010

Abstract

Background: The HUC-HEART Trial is a clinical study of intramyocardial delivery of current Good Manufacturing Practice (cGMP)-grade human umbilical cord multipotent stromal cells (HUC-MSCs) in ischemic cardiomyopathy where 2 × 10⁷ cells are administered to peri-infarcted myocardium. Prior to the onset of the trial, we aimed to optimize the transport/storage conditions for obtaining the highest cell viability and proliferation rate of cells to be transplanted.

Methods: Cells were tested after being transported in phosphate-buffered saline (PBS) or Ringer's lactate-based (RL) transport media supplemented with human serum albumin (HSA) and/or hydroxyethyl starch (HES) at two temperatures (2-10°C or 22-24°C).

Results: The effects of transport conditions on cell viability following 6 h were found highest (93.4 ± 1.5) in RL-based media at 2-10°C. Karyotypes were found normal upon transportation in any of the formulations and temperatures. However, the highest proliferation rate was noted (3.1-fold increase) in RL (1% HSA) media at 2-10°C over 6 days in culture. From that point, RL (1% HSA) media at 2-10°C was used for further experiments. The maximum cell storage time was detected around 24 h at 2-10°C. Extended storage periods resulted in a decrease in cell viability but not in MSC marker expression. An increase in actin quantity was detected in hypoxia (5% O₂) groups in early culture days; no difference was noted between hypoxic versus normoxic (21% O₂) conditions in later days.

Discussion: The overall results suggest that non-commercial, simple media formulations with extended storage intervals at 2-10°C temperatures are capable of retaining the characteristics of clinical-grade HUC-MSCs. The above findings led us to use RL (1% HSA) media at 2-10°C for transport and storage in the HUC-HEART Trial; 23 patients received HUC-MSCs by August 2018; no adverse effects were noted related to cell processing and transplantation.

Keywords: Wharton's jelly; cell storage; cell transport; clinical trial; mesenchymal stromal cell; umbilical cord.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actins / analysis
Cell Culture Techniques / methods*
Cell Hypoxia / physiology
Cell Proliferation
Cell Survival
Female
Humans
Infant, Newborn
Karyotype
Mesenchymal Stem Cell Transplantation*
Mesenchymal Stem Cells / physiology*
Myocardial Ischemia / therapy*
Specimen Handling / methods*
Temperature
Umbilical Cord / cytology*

Substances

Actins