Structural and Functional Characterization of Deceased Donor Stem Cells: A Viable Alternative to Living Donor Stem Cells

Abstract

Stem cells can be isolated from various human tissues including bone marrow (BM) and adipose tissue (AT). Our study outlines a process to isolate adult stem cells from deceased donors. We have shown that cell counts obtained from deceased donor BM were within established living donor parameters. Evaluation of demographic information exhibited a higher percentage of hematopoietic stem cells (HSC) in males versus females, as well as a higher percentage of HSC in the age bracket of 25 years and under. For the first time, we show that deceased donor femur BM grew cell colonies. Our introduction of new technology for nonenzymatic AT processing significantly increased cell recovery over the traditional enzymatic processing method. Cell counts from the deceased donor AT exceeded living donor parameters. Furthermore, our data illustrated that AT from female donors yielded a much higher number of total nucleated cells (TNC) than males. Together, our data demonstrates that our approach to isolate stem cells from deceased donors could be a routine practice to provide a viable alternative to living donor stem cells. This will offer increased accessibility for patients awaiting stem cell therapies.

Differentiation of Human Deceased Donor, Adipose-Derived, Mesenchymal Stem Cells into Functional Beta Cells

“…[23] , the amount of MSC found in adipose tissue is 1%-10% of total nucleated cells. In deceased donor adipose tissue, we have observed the MSC concentration to be 4.5% of the total nucleated cells [19] . The adipose-derived MSC were expanded in fresh media to yield cells that had spindle shape fibroblast-like morphology.…”

“…Adipose tissue may be procured on a routine basis from deceased donors during the normal deceased donor workup without the pain, morbidity and mortality associated with living donors. We have shown earlier that by using the adipose tissue obtained from deceased donors, we were able to isolate plastic-adherent MSC populations from the SVF having multi-lineage differentiation capacity [19] . As seen in Figure 1, flow cytometric characterization of MSC show that these cells express the stem cell-specific combination of surface markers such as CD73, CD90, CD29, CD44, and CD105 and the lack of expression of CD11b, CD45 [29] .…”

“…The isolated MSC from deceased donor adipose tissue maintained their differentiation capacity. After addition of appropriate growth factors, MSC could be differentiated to form adipocytes, chondrocytes and osteocytes as described earlier [19] .…”

“…Deceased donors are routinely HLA-typed and infectious disease screened. Adipose tissue can be easily procured from researchconsented deceased donors during the routine deceased donor workup without the pain, morbidity and mortality associated with living donor collection [19] . In the present study, we demonstrate that mesenchymal stem cells (MSC) isolated from deceased donor adipose tissue retain their stem cell characteristics and can be expanded to provide a ready source for cell differentiation studies.…”

Apheresis of Deceased Donors as a New Source of Mobilized Peripheral Blood Hematopoietic Stem Cells for Transplant Tolerance

“…Investigators at several academic centers have developed the ability to collect large numbers of HSCs from the bone marrow of deceased donors. This approach involves resection of a large portion of the donor spine, followed by a complex process to isolate stem cells from the crushed vertebrae 11,12 There are significant drawbacks to this technique, in that it is invasive and disfiguring, requires reconstruction of the spine for viewing the donor at a funeral home, necessitates additional surgical time in the operating room, there is a risk of contamination, and the process for isolating the HSCs is restricted to only a few highly specialized laboratories.…”