TY - JOUR
T1 - Fundamental Cryobiology of Human Hematopoietic Progenitor Cells I
T2 - Osmotic Characteristics and Volume Distribution
AU - Gao, D. Y.
AU - Chang, Q.
AU - Liu, C.
AU - Farris, K.
AU - Harvey, K.
AU - McGann, L. E.
AU - English, D.
AU - Jansen, J.
AU - Critser, J. K.
N1 - Funding Information:
1This work was supported by Methodist Hospital of Indiana, Inc.
Funding Information:
The authors thank Dr. David A. Williams for helpful discussions, Lawrence Mark for technical assistance, and K. Vernon for assistance in the preparation of the manuscript. This work is supported, in part, by the Methodist Hospital of Indiana.
PY - 1998/2
Y1 - 1998/2
N2 - While methods for the cryopreservation of hematopoietic stem cells are well established, new sources of progenitor cells, such as umbilical cord blood, fetal tissue, and ex vivo expanded progenitor cells, may require refined protocols to achieve optimal recovery after freezing. To predict optimal protocols for cryopreservation of human hematopoietic progenitors, knowledge of fundamental cryobiological characteristics including cell osmotic characteristics, water and cryoprotectant permeability coefficients of cell membrane, and activation energies of these coefficients is required. In this study, we used CD34+CD33- cells isolated from human bone marrow as hematopoietic progenitor cell models/representatives to study the osmotic characteristics of the progenitor cells. Volume distribution and osmotic behavior of the CD34+CD33- cells were determined using two different methods: (a) a shape-independent electronic sizing technique and (b) a shape-dependent optical image analysis. The cell diameter was measured to be 8.2 ± 1.1 μm (mean ± SD, n = 1,091,475, the number of donors = 8) using the electronic sizing technique or 8.7 ± 1.2 μm (mean ± SD, n = 1508, the number of donors = 6) by image analysis at initial (isotonic) osmolality, 325 mosm/kg. The cell volume change was measured after the cells were exposed and equilibrated to different anisosmotic conditions. The cell volume was found to be a linear function of the reciprocal of the extracellular osmolality (Boyle van't Hoff plot) ranging from 163 to 1505 mosm/kg. The volume fraction of intracellular water which is osmotically active was determined to be 79.5% of the cell volume. It was concluded that human CD34+CD33- cells osmotically behave as ideal osmometers. This information coupled with cell water and cryoprotectant permeability coefficients as well as their activation energies (to be determined in the ongoing research projects) will be used to design optimum conditions for cryopreservation of human hematopoietic progenitor cells.
AB - While methods for the cryopreservation of hematopoietic stem cells are well established, new sources of progenitor cells, such as umbilical cord blood, fetal tissue, and ex vivo expanded progenitor cells, may require refined protocols to achieve optimal recovery after freezing. To predict optimal protocols for cryopreservation of human hematopoietic progenitors, knowledge of fundamental cryobiological characteristics including cell osmotic characteristics, water and cryoprotectant permeability coefficients of cell membrane, and activation energies of these coefficients is required. In this study, we used CD34+CD33- cells isolated from human bone marrow as hematopoietic progenitor cell models/representatives to study the osmotic characteristics of the progenitor cells. Volume distribution and osmotic behavior of the CD34+CD33- cells were determined using two different methods: (a) a shape-independent electronic sizing technique and (b) a shape-dependent optical image analysis. The cell diameter was measured to be 8.2 ± 1.1 μm (mean ± SD, n = 1,091,475, the number of donors = 8) using the electronic sizing technique or 8.7 ± 1.2 μm (mean ± SD, n = 1508, the number of donors = 6) by image analysis at initial (isotonic) osmolality, 325 mosm/kg. The cell volume change was measured after the cells were exposed and equilibrated to different anisosmotic conditions. The cell volume was found to be a linear function of the reciprocal of the extracellular osmolality (Boyle van't Hoff plot) ranging from 163 to 1505 mosm/kg. The volume fraction of intracellular water which is osmotically active was determined to be 79.5% of the cell volume. It was concluded that human CD34+CD33- cells osmotically behave as ideal osmometers. This information coupled with cell water and cryoprotectant permeability coefficients as well as their activation energies (to be determined in the ongoing research projects) will be used to design optimum conditions for cryopreservation of human hematopoietic progenitor cells.
KW - Boyle van't Hoff plot
KW - CD34CD33 cells
KW - Human hematopoietic progenitors
KW - Osmotic behavior
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U2 - 10.1006/cryo.1997.2060
DO - 10.1006/cryo.1997.2060
M3 - Article
C2 - 9500931
AN - SCOPUS:0031989358
SN - 0011-2240
VL - 36
SP - 40
EP - 48
JO - Cryobiology
JF - Cryobiology
IS - 1
ER -