Studies on the bone marrow cells from an excised pipe bone.
Samples of rib bone were collected and preserved under various conditions. The effect of preserving a piece of bone on the yield of hematopoietic stem cells was analyzed. Bone marrow cells were collected by means of flushing the bone marrow cavity with medium. The initial number of nucleated bone marrow cells collected from the donor's excised rib ranged from 0.8 x 10(8) to 2.3 x 10(8) per 1 cm of rib with an average of 1.21 x 10(8) (n = 7). In addition, the initial number of mononuclear bone marrow cells ranged from 1.1 x 10(7) to 4.8 x 10(7) per 1 cm of rib with an average of 2.47 x 10(7) (n = 7). The percentage loss of mononuclear bone marrow cells from rib preserved in medium for 48 hours at 4 degrees C, was less than 30%. A colony-forming assay by the methylcellulose method, complemented with r-G-CSF, r-GM-CSF, r-IL3, r-IL6, and r-erythropoietin, was done serially using bone marrow buffy-coat cells. These cells were obtained from the pieces of rib that were stored under various conditions. The number of CFU-GM and BFU-E that proliferated in this culture was 41.1 +/- 5.5 and 15.2 +/- 2.7 per dish. If the bone was preserved at 4 degrees C for 48 hours following excision, the recovery of CFU-GM and BFU-E from the bone marrow cells was greater than 50%. The effect of cryopreserving bone marrow mononuclear cells on the yield of hematopoietic cells was also analyzed. The percentage loss of proliferated hematopoietic cells by cryopreservation was less than 30% for bone marrow cells. Bone marrow cells were collected from pieces of rib obtained from cadaveric donors by means of the same method. The average number of nucleated bone marrow cells and MNCs was 1.26 x 10(7) and 2.30 x 10(7) per 1 cm of rib, respectively. The average number of CFU-GM and BFU-E that was detected by the colony-forming assay was 33.4 +/- 5.35 and 11.7 +/- 1.75 per dish, respectively. The percentage loss of proliferated hematopoietic cells by cryopreservation was less than 40% for bone marrow cells from cadaveric donors. These data, which reveal that hematopoietic bone marrow stem cells could be collected from a piece of pipe bone that was without circulation, suggest that it is possible to use pipe bone from cadaveric donors as a source of hematopoietic stem cells for bone marrow transplants.(ABSTRACT TRUNCATED AT 400 WORDS)
Takakuwa
K
K
Department of Obstetrics and Gynecology, Niigata University School of Medicine, Japan.
Kurata
H
H
Hirono
T
T
Koike
T
T
Kishi
K
K
Shibata
A
A
Tanaka
K
K
D000368:Aged; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000379:methods; D002102:Cadaver; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods; D015925:Cryopreservation; D005260:Female; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D012272:Ribs / Q000166:cytology*; D013997:Time Factors