Phenotypic, Functional, and Safety Control at Preimplantation Phase of MSC‐Based Therapy

Abstract

Mesenchymal stem cells (MSC) exhibit enormous heterogeneity which can modify their regenerative properties and therefore influence therapeutic effectiveness as well as safety of these cells transplantation. In addition the high phenotypic plasticity of MSC population makes it enormously sensitive to any changes in environmental properties including fluctuation in oxygen concentration. We have shown here that lowering oxygen level far below air atmosphere has a beneficial impact on various parameters characteristic for umbilical cord Wharton Jelly- (WJ-) MSC and adipose tissue- (AD-) derived MSC cultures. This includes their cellular composition, rate of proliferation, and maintenance of stemness properties together with commitment to cell differentiation toward mesodermal and neural lineages. In addition, the culture genomic stability increased significantly during long-term cell passaging and eventually protected cells against spontaneous transformation. Also by comparing of two routinely used methods of MSCs isolation (mechanical versus enzymatic) we have found substantial divergence arising between cell culture properties increasing along the time of cultivation in vitro. Thus, in this paper we highlight the urgent necessity to develop the more sensitive and selective methods for prediction and control cells fate and functioning during the time of growth in vitro.

Comparison of the paracrine activity of mesenchymal stem cells derived from human umbilical cord, amniotic membrane and adipose tissue

“…Numerous studies have proven that cytokines produced by MSCs have an anti‐inflammatory effect . These properties can be very useful in the treatment of perinatal morbidities caused by excessive inflammatory response, such as respiratory distress, intracranial hemorrhage or infarction and necrotizing colitis …”

“…17,21 Numerous studies have proven that cytokines produced by MSCs have an anti-inflammatory effect. 16,[22][23][24][25][26] These properties can be very useful in the treatment of perinatal morbidities caused by excessive inflammatory response, such as respiratory distress, intracranial hemorrhage or infarction and necrotizing colitis. [10][11][12]27,28 Our results indicate that the secretion of cytokines by MSCs from various sources may be different.…”

Rescuing mesenchymal stem cell regenerative properties on hydrogel substrates post serial expansion

“…Eight‐arm 40 kDa PEG was functionalized with norbornene as previously described . Briefly, 5‐norbornene‐2‐carboxylic acid was coupled to eight‐arm 40 kDa PEG‐amine (Jenkem) in the presence of 1‐(bis[dimethylamino]methylene)‐1H‐1,2,3‐triazolo(4,5‐b)pyridinium3‐oxid hexafluorophosphate, N ‐([Dimethylamino]‐1H‐1,2,3‐triazolo‐[4,5‐b]pyridin‐1‐ylmethylene)‐ N ‐methylmethanaminium‐hexafluorophosphate N ‐oxide or HATU (Sigma) and N , N ‐diisopropylethylamine (Sigma), in dimethylformamide.…”

“…Eight-arm 40 kDa PEG was functionalized with norbornene as previously described. 29,30 Briefly, 5-norbornene-2-carboxylic acid was coupled to eight-arm 40 kDa PEG-amine (Jenkem) in the presence of…”

“…Peptide-functionalized PEG hydrogels were synthesized via a thiol-ene photoclick reaction. 29 Eight-arm PEG-thiols were co-polymerized with eight-arm PEG-norbornene (equal stoichiometry) to form predominantly elastic hydrogels (E = 1-20 kPa), where the final modulus was controlled by the concentration of PEG macromolecules in the initial solution (Figure 1a,b). The fibronectin-derived integrin binding motif, CRGDS, was incorporated into the hydrogels at a 2 mM concentration to promote hMSC attachment.…”