Abstract
Recent studies have demonstrated that the umbilical cord (UC) is an excellent source of mesenchymal stromal cells (MSCs). However, current protocols for extracting and culturing UC-MSCs do not meet current good manufacturing practice (cGMP) standards, in part due to the use of xenogeneic reagents. To support the development of a cGMP-compliant method, we have examined an enzyme-free isolation method utilizing tissue homogenization (t-H) followed by culture in human platelet lysate (PL) supplemented media. The yield and viability of cells after t-H were comparable to those obtained after collagenase digestion (Col-D). Importantly, kinetic analysis of cultured cells showed logarithmic growth over 10 tested passages, although the rate of cell division was lower for t-H as compared to Col-D. This slower growth of t-H-derived cells was also reflected in their longer population doubling time. Interestingly, there was no difference in the expression of mesenchymal markers and trilineage differentiation potential of cells generated using either method. Finally, t-H-derived cells had greater clonogenic potential compared to Col-D/FBS but not Col-D/PL and were able to maintain CFU-F capacity through P7. This bench scale study demonstrates the possibility of generating therapeutic doses of good quality UC-MSCs within a reasonable length of time using t-H and PL.
Highlights
A number of studies have highlighted the potential of mesenchymal stromal cells (MSCs) in tissue regeneration, immune regulation, and potentiation of ex vivo expansion of hematopoietic stem cells (HSCs) [1,2,3,4]
In order to disrupt the umbilical cord (UC) matrix and release the stromal cells, simple tissue homogenization (t-H) using the GentleMACS Dissociator was performed on 10 different UC samples
While enzymatic digestion results in a more uniform release of cells, it has the drawbacks of longer initial processing time, lack of a standard method, and being non-current good manufacturing practice (cGMP)-compliant since the reagents are obtained from nonhuman sources making it difficult to obtain clinical-grade material
Summary
A number of studies have highlighted the potential of mesenchymal stromal cells (MSCs) in tissue regeneration, immune regulation, and potentiation of ex vivo expansion of hematopoietic stem cells (HSCs) [1,2,3,4]. In vitro expanded UC-MSCs exhibit cell surface markers, differentiation capability, and immune regulatory properties comparable to those of BM-MSCs [7,8,9], with the added advantage of having a higher proliferation/expansion potential (greater numbers of passages to senescence) [8,9,10]. Such features reflect the relatively primitive nature of the UCMSCs compared to their adult counterpart. UCderived cells have not been exposed to viruses and toxins
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
