Clot-Derived Contaminants in Transplanted Bone Marrow Mononuclear Cells Impair the Therapeutic Effect in Stroke

Okinaka, Yuka; Kikuchi-Taura, Akie; Takeuchi, Kosei; Ogawa, Yuko; Boltze, Johannes; Gul, Sheraz; Claussen, Carsten; Taguchi, Akihiko

doi:10.1161/strokeaha.119.026669

Cited by 9 publications

(5 citation statements)

References 35 publications

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“…Furthermore, the results provide a potential explanation for the inconsistencies in the literature regarding the efficacy of BMC treatments for orthopedic conditions and RBC quantitation should be routinely reported in clinical studies evaluating BMC as a treatment option. Our findings have significant implications beyond the field of orthopedics and further the findings of others where RBC contamination poses deleterious effects on bone marrow mononuclear cell therapies following myocardial infarction and stroke [ 41 , 42 ]. Ultimately, clinicians should aim to produce BMC products with HCTs below 10% and HCT should be considered an indicator of product quality.…”

Section: Discussionsupporting

confidence: 78%

Red blood cells and their releasates compromise bone marrow-derived human mesenchymal stem/stromal cell survival in vitro

Dregalla¹,

Herrera²,

Donner

2021

Stem Cell Res Ther

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Purpose The use of bone marrow aspirate (BMA) and bone marrow aspirate concentrate (BMC) in the treatment of inflammatory orthopedic conditions has become a common practice. The therapeutic effect of BMA/BMC is thought to revolve primarily around the mesenchymal stem/stromal cell (MSC) population residing within the nucleated cell fraction. MSCs have the unique ability to respond to site of injury via the secretion of immunomodulating factors, resolving inflammation in diseased joints. Recently, the importance of hematocrit (HCT) in BMC has been debated, as the potential impact on MSC function is unknown. In the present study, we investigate MSC health over a short time-course following exposure to a range of HCT and red blood cell releasate (RBCrel) conditions. Methods Bone marrow-derived human MSCs in early passage were grown under conditions of 0%, 2.5%, 5%, 10%, 20% and 40% HCT and RBCrel conditions for 3 days. At each day, the percentage of viable, apoptotic and necrotic MSCs was determined via flow cytometry. Relative viable MSC counts in each condition was determined to account for dynamic changes in overall MSC densities over the time-course. Statistical analysis was performed using a one-way ANOVA comparing test conditions to the control followed by a Dunnett’s multiple comparison test. Results Significant reductions in viable MSCs concurrent with an increase in necrotic MSCs in high HCT and RBCrel conditions was observed within 24 h. At each successive timepoint, the percent and relative number of viable MSCs were reduced, becoming significant in multiple HCT and RBCrel conditions by Day 3. Necrosis appears to be the initial mode of MSC death following exposure to HCT and RBCrel, followed by apoptosis in surviving MSC fractions. Conclusion Various levels of HCT and RBCrel severely compromise MSC health within 3 days and HCT should be controlled in the preparation of BMC products. Further, HCT of BMCs should be routinely recorded and tracked with patient outcomes along with routine metrics (e.g. nucleated cell counts, fibroblast-colony forming units). Differences in HCT may account for the inconsistencies in the efficacy of BMC reported when treating orthopedic conditions.

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Section: Discussionsupporting

confidence: 78%

Red blood cells and their releasates compromise bone marrow-derived human mesenchymal stem/stromal cell survival in vitro

Dregalla¹,

Herrera²,

Donner

2021

Stem Cell Res Ther

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“…The femoral and tibial bones were dissected, distal ends were snipped using precision scissors, and BM was flushed out using PBS. BM was mechanically dissociated into single cells, and BM-MNCs were isolated by Ficoll-Paque (GE Healthcare, Little Chalfont, United Kingdom) density-gradient centrifugation as described elsewhere (Okinaka et al, 2019). Briefly, BM-MNCs were centrifuged at 400 × g for 40 min.…”

Section: Bm-mnc Preparationmentioning

confidence: 99%

Intravenous Bone Marrow Mononuclear Cells Transplantation in Aged Mice Increases Transcription of Glucose Transporter 1 and Na+/K+-ATPase at Hippocampus Followed by Restored Neurological Functions

Takeuchi

Okinaka

Ogawa

et al. 2020

Front. Aging Neurosci.

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We recently reported that intravenous bone marrow mononuclear cell (BM-MNC) transplantation in stroke improves neurological function through improvement of cerebral metabolism. Cerebral metabolism is known to diminish with aging, and the reduction of metabolism is one of the presumed causes of neurological decline in the elderly. We report herein that transcription of glucose transporters, monocarboxylate transporters, and Na + /K +-ATPase is downregulated in the hippocampus of aged mice with impaired neurological functions. Intravenous BM-MNC transplantation in aged mice stimulated the transcription of glucose transporter 1 and Na + /K +-ATPase α1 followed by restoration of neurological function. As glucose transporters and Na + /K +-ATPases are closely related to cerebral metabolism and neurological function, our data indicate that BM-MNC transplantation in aged mice has the potential to restore neurological function by activating transcription of glucose transporter and Na + /K +-ATPase. Furthermore, our data indicate that changes in transcription of glucose transporter and Na + /K +-ATPase could be surrogate biomarkers for age-related neurological impairment as well as quantifying the efficacy of therapies.

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“…Mice bone marrow mononuclear cells were prepared as we described previously (8). Briefly, bone marrow of the tibia and femur of 6 weeks old male C57BL/6N mice (Oriental yeast, Tokyo, Japan) was flushed out with phosphate buffered saline (PBS).…”

Section: Preparation Of Murine Bm-mncmentioning

confidence: 99%

Intravenous Bone Marrow Mononuclear Cells Transplantation Improves the Effect of Training in Chronic Stroke Mice

et al. 2020

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There is no effective treatment for chronic stroke if the acute or subacute phase is missed. Rehabilitation alone cannot easily achieve a dramatic recovery in function. In contrast to significant therapeutic effects of bone marrow mononuclear cells (BM-MNC) transplantation for acute stroke, mild and non-significant effects have been shown for chronic stroke. In this study, we have evaluated the effect of a combination of BM-MNC transplantation and neurological function training in chronic stroke. The effect of BM-MNC on neurological functional was tested four weeks after permanent middle cerebral artery occlusion (MCAO) insult in mice. BM-MNC (1 × 105cells in 100 μl PBS) were injected into the vein of MCAO model mice, followed by behavioral tests as functional evaluations. Interestingly, there was a significant therapeutic effect of BM-MNC only when repeated training was performed. This suggested that cell therapy alone was not sufficient for chronic stroke treatment; however, training with cell therapy was effective. The combination of these differently targeted therapies provided a significant benefit in the chronic stroke mouse model. Therefore, targeted cell therapy via BM-MNC transplantation with appropriate training presents a promising novel therapeutic option for patients in the chronic stroke period.

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Clot-Derived Contaminants in Transplanted Bone Marrow Mononuclear Cells Impair the Therapeutic Effect in Stroke

Cited by 9 publications

References 35 publications

Red blood cells and their releasates compromise bone marrow-derived human mesenchymal stem/stromal cell survival in vitro

Red blood cells and their releasates compromise bone marrow-derived human mesenchymal stem/stromal cell survival in vitro

Intravenous Bone Marrow Mononuclear Cells Transplantation in Aged Mice Increases Transcription of Glucose Transporter 1 and Na+/K+-ATPase at Hippocampus Followed by Restored Neurological Functions

Intravenous Bone Marrow Mononuclear Cells Transplantation Improves the Effect of Training in Chronic Stroke Mice

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