Skip to main content
SpringerLink
Log in

In vitro study on cryopreservation of peripheral blood stem cells with uncontrolled freezer

  • Published:
Chinese Journal of Cancer Research

Abstract

Objective: To evaluate the effects of cryopreservation of APBSCs in −80°C un-controlled freezer and liquid nitrogen, and to search for the efficient combined cryoprotectant and the highest cell concentration for cryopreservating peripheral blood stem cell (PBSC). Methods: To compare the effect of three combined cryoprotectants, evaluation of in vitro proliferative capacity by colony-forming unit-Granulocyte-macrophage (CFU-GM) and burst-forming Unit-erythroid (BFU-E) assay, immunophenotyping for CD34+/CD38− cells by FCM, and viability assessment by trypan blue exclusion were performed. Results: The cryoprotectant 10%DMSO+HES+auto-Plasma resulted in the highest recovery rates. CFU-GM and BFU-E were (78.7±9.8)%, (69.8±14.1)%, respectively. The recovery rates of CFU-GM and BFU-E in A/C groups were (68.3±6.2)%/ (65.8±7.2)% and (63.4±9.7)%/ (60.4±10.5)%, respectively. The recovery rates of CD34+/CD38− cells and cell viability with the three combined reagents were 90% and 80%, respectively. Conclusion: 5%DMSO+ HES+auto-PLASMA is an ideal combined cryoprotectant for cryopreserving PBSC. The cell concentrations may be cryopreserved at 4×108 ml−1.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Stiff PJ, Murgo AJ, Zaroulis CG, et al. Unfractionated human marrow cell cryopreservation using dimethylsulfoxide and hydroxyethyl starch[J]. Cryobiology 1983; 20: 17.

    Article  PubMed  CAS  Google Scholar 

  2. Donaldson C, Armitage WJ, Denning-kendall PA, et al. Optimal cryopreservation of human umbilical cord blood[J]. Bone Marrow Transplant 1996; 18: 725.

    PubMed  CAS  Google Scholar 

  3. Valeri CR, Pivacek LE. Effects of the temperature, the duration of frozen storage, and the freezing container on in vitro measurements in human peripheral blood mononuclear cells[J]. Transfusion 1996; 36: 303.

    Article  PubMed  CAS  Google Scholar 

  4. Galme’s A Besalduch J, Bargay J, et al. Long-term storage at −80°C of hematopoietic porgenitor cells with 5-percent dimethyl sulfoxide as the sole cryoprotectant[J]. Transfusion 1999; 39: 70.

    Article  CAS  Google Scholar 

  5. Luo KH, Wu GQ, Wang QL, et al. Effect of dimethylsulfoxide and hydroxyethyl starch in the preservation of fractionated human marrow cells[J]. Cryobiology 1994; 31: 349.

    Article  PubMed  CAS  Google Scholar 

  6. Luo KH, Shi YK, Sun Y, et al. A practical procedure for the cryopreservation of marrow cells intended for autolransplantatin[J]. Leuk and Lymphoma 1995; 17: 495.

    CAS  Google Scholar 

  7. Takaue Y, Abe T, Kawano Y, et al. Comparative analysis of engraftment after cryopreservation of PBSC autograft by controlled versus uncontrolled-rate methods[J]. Bone Marrow Transplant 1994; 13: 801.

    PubMed  CAS  Google Scholar 

  8. Katayama Y, Yano T, Bessho A, et al. The effects of a simplified method for cryopreservation and thawing procedures on peripheral blood stem cells[J]. Bone Marrow Transplant 1997; 19: 283.

    Article  PubMed  CAS  Google Scholar 

  9. Shlebak AA, Marley SB, Roberts IA, et al. Optimal timing for processing and cryopreservation of umbilical cord haematopoietic stem cells for clinical transplantation[J]. Bone Marrow Transplant 1999; 23: 131.

    Article  PubMed  CAS  Google Scholar 

  10. Humpe A, Riggert J, Vehmeyer K, et al. Comparison of CD34+ cell numbers and colony growth before and after cryopreservation of peripheral blood progenitor and stem cell harvests: influence of prior chemotherapy[J]. J Transfusion 1997; 37: 1050.

    Article  CAS  Google Scholar 

  11. Makino S, Harada M, Akashi K, et al. A simplified method for cryopreservation of peripheral blood stem cells at −80°C without rate-controlled freezing[J]. Bone Marrow Transplant 1991; 8: 239.

    PubMed  CAS  Google Scholar 

  12. Chen H. Collection and of Peripheral blood stem cell. In: Da Wan-ming and Pei Xue-tao, eds. Peripheral blood stem cell transplantation[M]. 1st ed. Beijing: People’s Health Publishing House, 2000; 91.

    Google Scholar 

  13. Rowley SD, Bensinger WI, Gooley TA, et al. Effect of cell concentration on bone marrow and peripheral blood stem cell cryopreservation[J]. Blood 1994; 83: 2731.

    PubMed  CAS  Google Scholar 

  14. Rowley SD. Recombinant human deoxyribonuclease for hematopoietic stem cell processing[J]. J hematother 1995; 4: 99.

    PubMed  CAS  Google Scholar 

  15. Xu YH. Blood. In: Zhang Jing-Ru, eds. Physiology[M]. 4th ed. Beijing: People’s Health Publishing House, 1996; 70.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Medical Oncology, Cancer Hospital (Institute), Chinese Academy of Medical Sciences and Peking Union Medical College, 100021, Beijing

    Zhang Jun-ping, Shi Yuan-kai, He Xiao-hui, Han Xiao-hong, Liu Peng & Yang Jian-liang

Authors
  1. Zhang Jun-ping
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shi Yuan-kai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. He Xiao-hui
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Han Xiao-hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Liu Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yang Jian-liang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shi Yuan-kai.

Additional information

Foundation item: This work was supported by a grant from the National “95” Key Project of China(No. 96-906-01-12).

Biography: Zhang Jun-ping(1965–), female, master of medicine, associate professor, now address: Shanxi Tumor Hospital, majors in hematological oncology.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, Jp., Shi, Yk., He, Xh. et al. In vitro study on cryopreservation of peripheral blood stem cells with uncontrolled freezer. Chin. J. Cancer Res. 14, 235–239 (2002). https://doi.org/10.1007/s11670-002-0052-1

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11670-002-0052-1

Key words

CLC number

Search

Navigation