Int J Stem Cells.  2018 Jun;11(1):26-38. 10.15283/ijsc17015.

CD73, CD90, CD105 and Cadherin-11 RT-PCR Screening for Mesenchymal Stem Cells from Cryopreserved Human Cord Tissue

Affiliations
  • 1Processing Laboratory, StemCyte International Cord Blood Therapeutic Company, Baldwin Park, CA, USA. mchen@stemcyte.com
  • 2Department of Medicine, University of California – Los Angeles, Los Angeles, CA, USA.
  • 3Department of Medicine, Veterans Affair, Greater Los Angeles Healthcare System, Los Angeles, CA, USA.
  • 4Tissue Laboratory, StemCyte, New Taipei City Linkou District, Taiwan.
  • 5University of Bologna, Sant' Orsola-Malpighi Hospital, Bologna, Italy.

Abstract

BACKGROUND AND OBJECTIVES
Mesenchymal stem cells (MSCs) are self-renewing, non-specialized cells used clinically in tissue regeneration and sourced from bone marrow, peripheral blood, umbilical cord blood and umbilical cord tissue (UCT). To demonstrate an alternative method for MSC detection, cryopreserved UCT and expanded MSC were screened for MSC markers CD73, CD90, CD105 and CDH-11 by RT-PCR.
METHODS AND RESULTS
Human UCT were washed, sectioned, cryopreserved with 10% DMSO and stored in the vapor phase of liquid nitrogen. Fresh and frozen UCT samples were expanded for MSC. UCT and MSC were processed for RNA and screened for CD73, CD90, CD105 and CDH-11 mRNA by RT-PCR. CD73, CD90 and CD105 were detected by flow cytometry and CDH-11 was detected by Western blotting. Short and long-term frozen UCT shows a loss of mRNA expression for CD73 at 33.2±34.0%, CD90 at 6.2±8.2%, CD105 at 17.7±21.6% and CDH-11 at 30.1±26.7% but was not statistically significant to indicate the deterioration. Expanded MSCs from fresh UCT expressed 0.09±0.07-fold CD73, 0.17±0.11-fold CD90, 0.04±0.06-fold CD105 and 0.14±0.08-fold CDH-11. Expanded MSCs from frozen UCTs expressed 0.09±0.06-fold CD73, 0.13±0.06-fold CD90, 0.04±0.05-fold CD105 and 0.11±0.06-fold CDH-11 and confirmed by flow cytometry and Western blotting.
CONCLUSION
CD73, CD90, CD105 and CDH-11 were detected by RT-PCR in cryopreserved UCT and MSC expansion. CDH-11 appears as a useful single target MSC marker for quick screening.

Keyword

CD73; CD90; CD105; Cadherin-11; Mesenchymal stem cells; Umbilical cord tissue
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