Large scale separation of human bone marrow by counterflow centrifugation elutriation

S. J. Noga; C. A. Cremo; S. C. Duff; C. L. Schwartz; A. Melaragno; C. I. Civin; A. D. Donnenberg

doi:10.1016/0022-1759(86)90168-7

Large scale separation of human bone marrow by counterflow centrifugation elutriation

S. J. Noga, C. A. Cremo, S. C. Duff, C. L. Schwartz, A. Melaragno, C. I. Civin, A. D. Donnenberg

Pediatrics

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Counterflow centrifugation elutriation (CCE) has been used to separate moderate quantities of bone marrow (BM) into distinct cell populations for further in vitro investigation. Recently, this technique was employed to reduce the incidence of graft-versus-host disease (GVHD) in human allogeneic bone marrow transplantation (BMT) by removing the majority of mature lymphocytes from the graft. Unfortunately, current methods employing the small (J6-B) elutriator rotor require time consuming preseparation steps and multiple runs. We report our experience with a fixed rotor speed, two-flow rate elutriation procedure using the new Beckman JE-10X rotor system which can separate more than 10¹⁰ nucleated BM cells in 30 min. Three fractions were obtained which differed in size, morphology, and immunologic capacity. The large cell fraction is suitable for allogeneic BMT since it is radically depleted of mature T lymphocytes and retains high clonogeneic capacity.

Original language	English (US)
Pages (from-to)	211-218
Number of pages	8
Journal	Journal of Immunological Methods
Volume	92
Issue number	2
DOIs	https://doi.org/10.1016/0022-1759(86)90168-7
State	Published - Sep 27 1986

Keywords

Beckman JE-10X rotor
Bone marrow, human
Colony-forming assay
Counterflow centrifugation elutriation
Mixed lymphocyte response
Phytohemagglutinin assay

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/0022-1759(86)90168-7

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title = "Large scale separation of human bone marrow by counterflow centrifugation elutriation",

abstract = "Counterflow centrifugation elutriation (CCE) has been used to separate moderate quantities of bone marrow (BM) into distinct cell populations for further in vitro investigation. Recently, this technique was employed to reduce the incidence of graft-versus-host disease (GVHD) in human allogeneic bone marrow transplantation (BMT) by removing the majority of mature lymphocytes from the graft. Unfortunately, current methods employing the small (J6-B) elutriator rotor require time consuming preseparation steps and multiple runs. We report our experience with a fixed rotor speed, two-flow rate elutriation procedure using the new Beckman JE-10X rotor system which can separate more than 1010 nucleated BM cells in 30 min. Three fractions were obtained which differed in size, morphology, and immunologic capacity. The large cell fraction is suitable for allogeneic BMT since it is radically depleted of mature T lymphocytes and retains high clonogeneic capacity.",

keywords = "Beckman JE-10X rotor, Bone marrow, human, Colony-forming assay, Counterflow centrifugation elutriation, Mixed lymphocyte response, Phytohemagglutinin assay",

author = "Noga, {S. J.} and Cremo, {C. A.} and Duff, {S. C.} and Schwartz, {C. L.} and A. Melaragno and Civin, {C. I.} and Donnenberg, {A. D.}",

note = "Funding Information: This work was supported by Grants CA15396, CA31687, CA3218, CA06973, CA09071 from the National Institutes of Health, work unit number M0095.001.0003 from NMR and DC, and generous gifts from the Royal Sisters Society of Baltimore, MD, Beckman Instruments, Palo Alto, CA and Beckton-Dickinson, Mountainview, CA. Dr. Civin is a scholar of the Leukemia Society of America.",

year = "1986",

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doi = "10.1016/0022-1759(86)90168-7",

language = "English (US)",

volume = "92",

pages = "211--218",

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T1 - Large scale separation of human bone marrow by counterflow centrifugation elutriation

AU - Noga, S. J.

AU - Cremo, C. A.

AU - Duff, S. C.

AU - Schwartz, C. L.

AU - Melaragno, A.

AU - Civin, C. I.

AU - Donnenberg, A. D.

N1 - Funding Information: This work was supported by Grants CA15396, CA31687, CA3218, CA06973, CA09071 from the National Institutes of Health, work unit number M0095.001.0003 from NMR and DC, and generous gifts from the Royal Sisters Society of Baltimore, MD, Beckman Instruments, Palo Alto, CA and Beckton-Dickinson, Mountainview, CA. Dr. Civin is a scholar of the Leukemia Society of America.

PY - 1986/9/27

Y1 - 1986/9/27

N2 - Counterflow centrifugation elutriation (CCE) has been used to separate moderate quantities of bone marrow (BM) into distinct cell populations for further in vitro investigation. Recently, this technique was employed to reduce the incidence of graft-versus-host disease (GVHD) in human allogeneic bone marrow transplantation (BMT) by removing the majority of mature lymphocytes from the graft. Unfortunately, current methods employing the small (J6-B) elutriator rotor require time consuming preseparation steps and multiple runs. We report our experience with a fixed rotor speed, two-flow rate elutriation procedure using the new Beckman JE-10X rotor system which can separate more than 1010 nucleated BM cells in 30 min. Three fractions were obtained which differed in size, morphology, and immunologic capacity. The large cell fraction is suitable for allogeneic BMT since it is radically depleted of mature T lymphocytes and retains high clonogeneic capacity.

AB - Counterflow centrifugation elutriation (CCE) has been used to separate moderate quantities of bone marrow (BM) into distinct cell populations for further in vitro investigation. Recently, this technique was employed to reduce the incidence of graft-versus-host disease (GVHD) in human allogeneic bone marrow transplantation (BMT) by removing the majority of mature lymphocytes from the graft. Unfortunately, current methods employing the small (J6-B) elutriator rotor require time consuming preseparation steps and multiple runs. We report our experience with a fixed rotor speed, two-flow rate elutriation procedure using the new Beckman JE-10X rotor system which can separate more than 1010 nucleated BM cells in 30 min. Three fractions were obtained which differed in size, morphology, and immunologic capacity. The large cell fraction is suitable for allogeneic BMT since it is radically depleted of mature T lymphocytes and retains high clonogeneic capacity.

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KW - Phytohemagglutinin assay

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Large scale separation of human bone marrow by counterflow centrifugation elutriation

Abstract

Keywords

ASJC Scopus subject areas

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