Effects of cryopreservation and long-term culture on biological characteristics and proteomic profiles of human umbilical cord-derived mesenchymal stem cells

Abstract

Background: Human umbilical cord-derived MSCs (hUC-MSCs) have been identified as promising seeding cells in tissue engineering and clinical applications of regenerative medicine due to their advantages of simple acquisition procedure and the capability to come from a young tissue donor over the other MSCs sources. In clinical applications, large scale production is required and optimal cryopreservation and culture conditions are essential to autologous and allogeneic transplantation in the future. However, the influence of cryopreserved post-thaw and long-term culture on hUC-MSCs remains unknown, especially in terms of specific protein expression. Therefore, biological characteristics and proteomic profiles of hUC-MSCs after cryopreserving and long-term culturing were investigated. Methods: Firstly, hUC-MSCs were isolated from human umbilical cord tissues and identified through morphology, surface markers and tri-lineage differentiation potential at passage 3, and then the biological characteristics and proteomic profiles were detected and compared after cryopreserving and long-term culturing at passage 4 and continuously cultured to passage 10 with detection occurring here as well. The proteomic profiles were tested by using the isobaric tags for relative and absolute quantification (iTRAQ) labeling technique and differential protein were confirmed by mass spectrometry. Results: The results showed no significant differences in phenotypes including morphology, surface marker and tri-lineage differentiation potential but have obvious changes in translation level, which is involved in metabolism, cell cycle and other pathways. Conclusion: This suggests that protein expression may be used as an indicator of hUC-MSCs security testing before applying in clinical settings, and it is also expected to provide the foundation or standardization guide of hUC-MSCs applications in regenerative medicine.

Assessment of Long-Term in vitro Multiplied Human Wharton’s Jelly-Derived Mesenchymal Stem Cells prior to Their Use in Clinical Administration

“…In our study, cultured cells were confirmed as WJ-MSCs by immunophenotyping, where they have expressed MSC surface markers CD90 greater than 90%, while CD105 expression was below 90%, which was not according to the ISCT guidelines. In a recent study done by Fu et al 2020], their cultured human umbilical cord-derived MSCs have shown expression of CD105 of 85.0 ± 1.4%, while we have obtained expression of CD105 of 88.89%, though the study by Pham and his group [2019] has suggested that there is the possibility of an unsettled expression of positive marker CD105, where even CD105-negative umbilical cord MSCs have shown 3.3% positive expression of CD105, which have been characterized as MSCs.…”

Thawed cryopreserved synovial mesenchymal stem cells show comparable effects to cultured cells in the inhibition of osteoarthritis progression in rats

“…However, reports on the influence of cryopreservation of MSCs on gene and protein expression are limited. Fu et al showed that cryopreservation of human umbilical cord-derived MSCs affected the expressions of proteins related to metabolism and cell cycle pathways 35 . We need to clarify whether cryopreservation of synovial MSCs affects the expression of proteins related to cell survival, anti-inflammation, and chondrogenesis.…”

Mesenchymal stromal cell therapy for coronavirus disease 2019: which? when? and how much?

“…To achieve bedside transplantation, a common practice is to thaw the vial containing a predetermined dose, resuspend in an electrolyte solution and inject. While this method is logistically convenient, allows rapid access and availability and eases the process of quality control [ 94 , 95 ], it can severely hamper viability if not done quickly by experienced handlers. While the majority of literature suggests minimal to no adverse effect of cryopreservation on MSC morphology, immunophenotype, proliferation and trilineage differentiation [ 95 , 96 ], very few studies have investigated immunomodulatory parameters.…”

“…While this method is logistically convenient, allows rapid access and availability and eases the process of quality control [ 94 , 95 ], it can severely hamper viability if not done quickly by experienced handlers. While the majority of literature suggests minimal to no adverse effect of cryopreservation on MSC morphology, immunophenotype, proliferation and trilineage differentiation [ 95 , 96 ], very few studies have investigated immunomodulatory parameters. Moll et al demonstrated a significant increase in IBMIR after blood exposure of early passage (P2-P4) thawed MSCs as opposed to fresh cultures in addition to increased complement-mediated cell lysis.…”

“…While the majority of literature suggests minimal to no adverse effect of cryopreservation on MSC morphology, immunophenotype, proliferation and trilineage differentiation (95,96), very few studies have investigated immunomodulatory parameters. Moll et al demonstrated a significant increase in IBMIR after blood exposure of early passage (P2-P4) thawed MSCs as opposed to fresh cultures in addition to increased complement-mediated cell lysis.…”