Manufacturing of Wharton’s Jelly -Derived MesenchymalS tem Cells (WJ-MSCs) by Enzymatic Cocktail

Objective: The isolation of Wharton’s Jelly -Derived MesenchymalS tem Cells (WJ-MSCs) from naïve umbilical cord tissue(UC) is particularly important for clinical use; therefore,according to the increasing demand for cellular therapy, it needsto be safe, cos t effective, and scaled up. However, current methodsto isolate MSCs from WJ yield low amounts of cells withvariable proliferation potentials. In this s tudy we successfullydesigned a new protocol to derivate MSCs by enzymatic approachfrom fresh samples which presents efficiency and consistency as it allows simultaneous processing of multiple UCsamples to produce confluent cultures in a predictable and shorttimeframe, without impacting on many of the properties that arecharacteris tic of MSCs or potentially therapeutic.Materials and Methods: UC tissue was cut into smaller pieces.Minced tissue was diges ted with collagenase I at ۳۷°C andthen, centrifuged at ۴۰۰ g. Supernatant was removed and theenzymatic cocktail (collagenase I and hyaluronidase) added tothe pellet for further diges tion, and incubated for about ۲۰ minutes.Finally, the enzyme s top solution was added to terminatethe incubation period; then, filtrated by cell s trainer.Results: The enzymatic cocktail can be used to isolate largenumbers of MSCs in less than ۲۴ hours. All the fresh isolatedMSCs had typical fibroblas tic morphology, expressed MSCsurfacemarkers (CD۷۳, CD۹۰, CD۱۰۵), and effectively differentiatedinto three mesodermal lineages. The cell count pergram of WJ processed by enzymatic method was ۴.۰۸ ± ۱.۲۵ ×۱۰۵ cells, after ۵ days of primary culture (p۰); which is significantlyon average ۱.۴ higher when compared with explantationapproach for the same WJ, but after ۲۵ days of p۰ (۳.۳ ± ۰.۷۷×۱۰۵ cells).Conclusion: By this protocol, we successfully reduced the requiredtime of primary culture to produce large-amount andhigh-quality MSCs, which will be suitable for cord tissue banksin the case of providing therapeutic services.

کلیدواژه ها:

Enzymatic Cocktail ، Isolation ، Mesenchymal S tem Cells ، Umbilical Cord Tissue ، Wharton’s Jelly