Table 7 The induction of apoptosis in post-thaw BM-MSCs

Apoptosis

 Human

  Liu et al. [28]

Human

Serum-free reduced-DMSO freezing solution gives comparable apoptotic percentage to 10% DMSO

Flow cytometry

  Ginis et al. [50]

Human

Lower percentage of apoptotic cells obtained with Annexin V and Hoechst staining compared to caspase 3 assay: using caspase 3, the percentage of apoptotic cells was between 13 and 17% for CryStor media compared to 3% for conventional freezing media

Flow cytometry—Annexin V, Hoechst, Caspase 3 activity

  Chinnadurai et al. [20]

Human

Higher percentage of apoptotic cells in cryopreserved MSC than live MSC

Flow cytometry

  Moll et al. [38]

Human

Apoptosis increased by cryopreservation when exposed to human serum

Flow cytometry—Annexin V, PI staining

 Other species

  Ock and Rho [51]

Pig

Bak and Bcl2 gene expression in cryopreserved cells was higher than fresh at 3 h post-thaw: Bak and Bcl2 gene expression in cryopreserved cells was comparable to fresh after culturing thawed cells up to 90% confluence: Bcl2 antigen expression level was comparable to fresh after culturing thawed cells up to 90% confluence

RT-qPCR for Bak and Bcl2: Flow cytometry; Bcl2 antigen

  Romanek et al. [98]

Pig (BM-MSC treated with HHP before freezing)

No significant difference between control (without HHP) and cells subjected to HHP pre-freeze

Flow cytometry—Annexin V: Fluorescence microscopy

Senescence

 Human

  Mamidi et al. [33]

Human

No difference in the level of senescent cells

Β-galactosidase assay

  Al-Saqi et al. [66]

Human

There were signs of senescence (but could be due to culture medium rather than cryopreservation medium)

Β-galactosidase assay (analysed 2 passages after cryopreservation)

  Pollock et al. [67]

Human

Immediate pre-freeze senescence levels show similar trends but higher levels compared to pre-freeze At 48 h post-thaw, level of senescent cells dropped significantly comparing to immediately post-thaw

Beta-glo assay