Concentration of rat bone marrow nucleated cells using hypo-osmotic hemolysis in distilled water

Fumio Ohnishi, Hiroko Ochiai, Kazuo Kishi

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Bone marrow aspirates contain primarily red blood cells. To achieve efficient cell transplantation for regeneration, the red blood cells need to be removed from the aspirates. Cell isolation is typically performed using density gradient centrifugation. However, this method entails issues of clinical safety and convenience. This study describes an efficient method to concentrate bone marrow nucleated cells by hypo-osmotic hemolysis. METHODS: The optimal hemolysis conditions were determined by diluting the bone marrow suspensions with distilled water in various dilution ratios. Then, the resulting cell fractions were transplanted in a rat cranial defect model to evaluate their effects on bone formation and their angiogenic effects. RESULTS: The optimal hemolysis conditions were a 3.3-fold dilution in distilled water and a hypo-osmotic exposure time of 45 seconds. Nucleated cells obtained using this method included granulocytes and mononuclear cells. These cells contain cytoplasmic angiogenic factors, including vascular endothelial growth factor, basic fibroblast growth factor, and hepatocyte growth factor. In a rat cranial defect model, callus formation and angiogenesis were significantly increased following transplantation of concentrated marrow nucleated cells in this manner. CONCLUSIONS: These results suggest angiogenic and osteogenetic effects of transplanting marrow nucleated cells using this hypo-osmotic method.This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License, where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially. http://creativecommons.org/licenses/by-nc-nd/3.0. American Society of Plastic Surgeons.

Original languageEnglish
JournalPlastic and Reconstructive Surgery
DOIs
Publication statusAccepted/In press - 2014 Jul 22

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Hemolysis
Bone Marrow Cells
Water
Bone Marrow
Licensure
Erythrocytes
Hepatocyte Growth Factor
Density Gradient Centrifugation
Cell Separation
Angiogenesis Inducing Agents
Cell Transplantation
Bony Callus
Fibroblast Growth Factor 2
Granulocytes
Osteogenesis
Vascular Endothelial Growth Factor A
Regeneration
Suspensions
Transplantation
Safety

ASJC Scopus subject areas

  • Surgery

Cite this

Concentration of rat bone marrow nucleated cells using hypo-osmotic hemolysis in distilled water. / Ohnishi, Fumio; Ochiai, Hiroko; Kishi, Kazuo.

In: Plastic and Reconstructive Surgery, 22.07.2014.

Research output: Contribution to journalArticle

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abstract = "BACKGROUND: Bone marrow aspirates contain primarily red blood cells. To achieve efficient cell transplantation for regeneration, the red blood cells need to be removed from the aspirates. Cell isolation is typically performed using density gradient centrifugation. However, this method entails issues of clinical safety and convenience. This study describes an efficient method to concentrate bone marrow nucleated cells by hypo-osmotic hemolysis. METHODS: The optimal hemolysis conditions were determined by diluting the bone marrow suspensions with distilled water in various dilution ratios. Then, the resulting cell fractions were transplanted in a rat cranial defect model to evaluate their effects on bone formation and their angiogenic effects. RESULTS: The optimal hemolysis conditions were a 3.3-fold dilution in distilled water and a hypo-osmotic exposure time of 45 seconds. Nucleated cells obtained using this method included granulocytes and mononuclear cells. These cells contain cytoplasmic angiogenic factors, including vascular endothelial growth factor, basic fibroblast growth factor, and hepatocyte growth factor. In a rat cranial defect model, callus formation and angiogenesis were significantly increased following transplantation of concentrated marrow nucleated cells in this manner. CONCLUSIONS: These results suggest angiogenic and osteogenetic effects of transplanting marrow nucleated cells using this hypo-osmotic method.This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License, where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially. http://creativecommons.org/licenses/by-nc-nd/3.0. American Society of Plastic Surgeons.",
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