Femur bone marrow from brain death deceased donors as source of human mesenchymal stromal cells for cell therapy

脑死亡尸体捐献者的股骨骨髓作为细胞治疗的人类间充质基质细胞的来源

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作者:Lourdes Echarte, Alexandra Sujanov, Daniel Machin, Natalia Marquisá, Cristina Touriño

Background

The use of a deceased donor (DD) as an alternative source of human mesenchymal stromal cells (hMSC) is promising, but has been little explored. This study evaluated the potential of femur bone marrow (FBM) from brain-death donors as a source of hMSC and compared this with hMSC from matched iliac crest bone marrow (ICBM).

Conclusions

To our knowledge, there are no previous studies evaluating BM from femur dead donors as a source of hMSC. Our findings confirm that it is feasible to expand cells from FBM from brain-death donors meeting in vitro characteristics of hMSC, making them a promising source for clinical translation.

Methods

Sixteen donor-matched FBM and ICBM samples were processed from brain-death donors. We analyzed the starting material and compared cell yield, phenotypic profile and differentiation capacity of hMSC.

Results

Neither the amount of nucleated cells per gram (14.6×106±10.3×106 from FBM vs. 38.8×106±34.6×106 from ICBM, P≥0.09) nor the frequency of CFU-F (0.0042%±0.0036% in FBM vs. 0.0057%±0.0042% in ICBM, P≥0.73) differ significantly from FBM or ICBM. Cell cultures from both sources were obtained and hMSC yields showed that there were no significant differences in hMSC obtained per gram of bone marrow (BM) when comparing femur with iliac crest samples. At passage 2, 12.5×106±12.9×106 and 5.0×106±4.4×106 hMSC per gram of BM were obtained from FBM and ICBM, respectively. FBM and ICBM hMSC express CD73, CD90, CD105, but not hematopoietic lineage markers [CD45, CD34, CD11, CD19 and isotype of HLA clase II (HLA-DR)]. HLA-A expression from both sources was clearly detected, while HLA-B was weakly expressed or undetectable and HLA-DR was undetectable. Cells from both sources were differentiated in vitro into osteoblasts, adipocytes and chondroblasts. Conclusions: To our knowledge, there are no previous studies evaluating BM from femur dead donors as a source of hMSC. Our findings confirm that it is feasible to expand cells from FBM from brain-death donors meeting in vitro characteristics of hMSC, making them a promising source for clinical translation.

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