Long-Term Cryopreservation: Successful Trilineage Engraftment After Autologous Bone Marrow Transplantation with Bone Marrow Cryopreserved for Seven Years

Abstract

Successful autologous bone marrow transplantation (ABMT) and peripheral blood stem cell transplantation depend on safe hematopoietic stem cell (HSC) cryopreservation and storage. Several successful methods for cryopreservation and storage have been established and are commonly used all over the world. However, little is known about the effects of long-term cryopreservation on the capacity to sustain a complete immunohematopoietic engraftment. Several authors have investigated stem cell viability after cryopreservation and storage for more than 5 years and reported preclinical good viabilities in terms of dye-exclusion or colony-forming capability in vitro. Only two studies using BM cryopreserved for more than 5 years for transplantation are reported, but they did not provide proof of trilineage engraftment. In February 1997 at our institution, a patient with relapsed non-Hodgkin's lymphoma underwent ABMT with BM harvested in February 1990. He achieved a granulocyte count > 500 x 10(6)/L on day 21 and a self-supporting platelet count > 20 x 10(9)/L on day 30. After day 29, his hemoglobin level was satisfactory without need of transfusion support. This successful trilineage engraftment with cryopreserved BM that had been stored for 7 years suggests that HSC are able to maintain their capability to reconstitute hematopoiesis for a long time.

Cryopreservation of Human Hematopoietic Cells with Membrane Stabilizers and Bioantioxidants as Additives in the Conventional Freezing Medium

“…However, cryopreservation at 280°C is a need in transplantation centers where rate-controlled and liquid nitrogen freezing facilities are not available due to financial constraints (31)(32)(33). Lastly, long-term freezing is particularly important for CB and fetal hematopoietic cells because the need to revive these tissues may occur months to years after freezing, depending on the demand for a matched tissue (34,35). Our results are encouraging because trehalose was found to exert its cryoprotective effect under all the three situations, i.e., storage at ultra-low temperatures or at 280°C and storage for extended periods.…”

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“…Cells are frozen at high cell density (20-40 million cells ml −1 ) in cryo-bags, previously used successfully in blood cell banking (Regidor et al, 1999). Frozen cell bag stability has been reported for more than 7 yr when used to store umbilical cord blood (Re et al, 1998), thus suggesting the possibility of using these bags for mammalian cell lines producing recombinant proteins. The cells are thawed and directly transferred into a dedicated 'inoculation' reactor that serves as the seed source for production scale campaigns.…”

The emergence of cellular therapy: impact on transfusion medicine

“…The practicality of stem cell product disposal raises a variety of administrative, clinical, and ethical issues that have previously never been addressed by blood bank staff. The difficult scenarios include: 1) the patient is not known to be deceased, but cannot otherwise be located; 2) although the patient is still alive the oncologist believes it is unlikely that the stored cells will ever be used; 3) the product was collected over a decade ago, and while theoretically, frozen stems cells could be stored indefinitely, there are little data studying frozen stored cell viability in humans over long storage periods; 15 4) a patient may relapse at some point after any given arbitrary storage period when the product has already been thrown out. A very difficult scenario at our institution was when the parents of a deceased child were contacted about stem cells that had been collected.…”