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Table 5 Experimental studies evaluating post-thaw BM-MSCs proliferation potential

From: The impact of cryopreservation on bone marrow-derived mesenchymal stem cells: a systematic review

Study

Species

Results post-thaw

Method of assessment

Human

 Bruder et al. [91]

Human

No effect on proliferation

Cell re-plated for one passage post-thaw; crystal violet dye-binding method

 Haack-Sorensen et al. [19]

Human

No effect on proliferation

PKH26-GL cell linker kit

 Xiang et al. [93]

Human

No effect on proliferation

Growth curves

 Zhao et al. [94]

Human (with chronic myeloid leukaemia)

No effect on proliferation

Cell count and cell-doubling time

 Doan et al. [95]

Human

No effect on proliferation

NA

 Ginis et al. [50]

Human

Proliferation of cryopreserved cells after 1 or 5 months of storage was higher than non-cryopreserved cells

Calcein-AM staining (at day 1, 4, 7 and 14 after post-thaw plating)

 Mamidi et al. [33]

Human

No effect on proliferation

Population doublings, cumulative population doublings and population doubling time

 Matsumura et al. [26]

Human

No effect on proliferation

Cell count; population doubling time (24 h, 48 h, 72 h and 96 h post-thaw)

 Holubova et al. [69]

Human

No effect on proliferation

Cell count

 Al-Saqi et al. [66]

Human

No significant difference in population doubling time but cells cryopreserved in DMSO had longer population doubling time compared to fresh

Population doubling (first and second passage post-thaw)

 Luetzkendorf et al. [40]

Human

No effect on proliferation

Population doublings; Population doubling time

 Pollock et al. [67]

Human

Population doublings decreased with increasing pre-freeze passage number

Population doublings

 Lechanteur et al. [34]

Human

Very low recovery until day 4 then a slight increase indicating re-proliferation

Cell count (0–5 days after thawing)

 Yuan et al. [52]

Human (BM-MSC engineered to express TRAIL)

No effect on proliferation

XTT assay

Other species

 Edamura et al. [36]

Dog

DMSO and FBS-free freezing resulted in similar proliferative capacity as non-cryopreserved; DMSO and FBS containing freezing media gave lower proliferative capacity

Cell count (2, 4, 6,8, 10 and 12 days post-thaw)

 Tokumoto et al. [48]

Monkey

No effect on proliferation

DNA quantification at 4, 8 and 12 days

 Lauterboeck et al. [49]

Monkey

No effect on proliferation

Population doubling time

 Heino et al. [39]

Minipig

Two to sixfolds decrease in the proliferative capacity of cells

Population doublings

 Romanek et al. [98]

Pig (BM-MSC treated with a high hydrostatic pressure (HHP) before freezing)

Cells treated with HHP showed better proliferation rate

Cell count

 Mitchell et al. [32]

Horse

No effect on proliferation

Cell staining with CellTrace label

Colony-forming unit ability

 Human

  Verdanova et al. [25]

Human

Best number of colonies obtained when cells were frozen with 5% DMSO with 5% sericin in culture medium

Cells seeded 60 cm Petri dishes for 2 weeks, Crystal Violet stained and colonies counted (light microscope)

 Other species

  Ock and Rho, [51]

Pig

All cryopreserved cells showed significantly lower numbers of colonies compared to fresh; Lower DMSO produced higher number of colonies

Cells seeded in 6-well plates for 2 weeks, 4% Giemsa stained and colonies counted (light microscope)

  Mitchell et al. [32]

Horse

No effect on colony-forming unit ability

Cells seeded in 10 cm plates for 1 week, Crystal Violet stained and colonies counted (light microscope)

  1. The key results on bone-marrow derived mesenchymal stem cell proliferation are presented in this table. For further details on the cryopreservation experimental details refer to either Table 1 or Additional file 2 which provide the individual freezing protocols outlined in the extracted papers alongside the concentration and passage of cells at the point of cryopreservation and the process of thawing