Characterization of cryopreserved human Langerhans cells.
Epidermal Langerhans cells are potent antigen-presenting cells in the epidermis. The establishment of a cryopreservation method for human Langerhans cells would greatly contribute to our ability to successfully conduct various experiments dealing with Langerhans cells. Since Langerhans cells are known to be sensitive to cold injury, there have been no reports concerning the cryopreservation of Langerhans cells. We have investigated the effect of cryopreservation on the function and phenotype of human Langerhans cells. Langerhans cells from human foreskins were isolated with the immunomagnetic microbead method using monoclonal antibodies for CD1a. Langerhans cells were cryopreserved in the presence of dimethylsulfoxide (DMSO) 10% and fetal calf serum 90%. Cryopreserved Langerhans cells were phenotypically assessed by flowcytometry using monoclonal antibodies to HLA-DR and CD1a. The ultrastructures of the Langerhans cells were compared using electron microscopy. An autologous T cell stimulation test was performed to compare the functions of cryopreserved Langerhans cells and fresh Langerhans cells. The viability of the cryopreserved Langerhans cells was able to be maintained at more than 90%. Cryopreserved Langerhans cells expressed high levels of HLA-DR and CD1a antigens and stimulated autologous T cells to an extent almost identical to that obtained from fresh Langerhans cells. These findings indicate that the cryopreservation of human Langerhans cells could lead to a breakthrough in various experiments dealing with human Langerhans cells.
Seo
Koo Il
KI
Department of Dermatology, Seoul National University College of Medicine, 28, Yungon-Dong, Chongno-Gu, Seoul 110-744, Republic of Korea.
Huh
Chang Hun
CH
Han
Ji Hyun
JH
Youn
Jai Il
JI
Lee
Chul Hee
CH
Lee
Wang Jae
WJ
Eun
Hee Chul
HC
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1 / Q000378:metabolism; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D016130:Immunophenotyping; D066298:In Vitro Techniques; D007801:Langerhans Cells / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D008213:Lymphocyte Activation; D008854:Microscopy, Electron; D013601:T-Lymphocytes / Q000276:immunology; D013936:Thymidine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1; C419393:CD1a antigen; D003451:Cryoprotective Agents; D006684:HLA-DR Antigens; D013936:Thymidine; D004121:Dimethyl Sulfoxide