Cord blood (CB)stem cells for wound repair
Preliminary report of 2cas
M.Valbonesi
a,*
,G.Giannini a ,F.Migliori b ,R.Dalla Costa b ,A.M.Dejana
a
a
Immunohematology Services,Department of Immunohematology,San Martino University Hospital,Largo Rosanna Benzi 10,ashlynn
16132Genova,Italy
b
savage garden
Immunohematology Services,Department of Immunohematology,Plastic Surgery Division,San Martino University Hospital,
Largo Rosanna Benzi 10,16132Genova,Italy
Accepted 26November 2003
whenever是什么意思Abstract
In 2patients,to promote skin wound/lesion repair we ud fibrin-platelet glue combined with HLA compatible (2mismatches accepted)buffy coats containingCD 34+cord blood cells.
The fibrin platelet glue was prepared with autologous apheresis platelets and cryoprecipitate.The original product was divided into 3and 4aliquots respectively for a correspondent number of applications.At each application,the margins of the lesion were infiltrated with 3ml of cord blood buffy coat,containing 30·103CD 34+cells.No graft versus tissue reaction was en in our patients in a follow-up of 3–7months.
The level of improvement,scored arbitrarily from 0to 4,was 3and 4,respectively.Our conclusion is that the u of cord blood cells alongwith fibrin platelet g lue is of clinical interest.Ó2004Elvier Ltd.All ri
ghts rerved.
Keywords:Cord blood;Progenitors stem cells;Wound repair;Platelet glue;Fibrin glue
1.Introduction
The clinical u of CB dates back to 1989when the first CB transplant was successfully carried out in a patient with Fanconi anemia [1].Since then CB has also been ud with some success for unrelated transplantation usingboth H LA identi-cal but also mismatched units [2].Conquently,cord blood bank systems have been t up both in
Europe and in the States [3,4]and CB cells are employed for transplantation both becau of their ready availability for patients lackingrelated and unrelated HLA identical donor and becau of the reduced risk of vere acute or chronic GVHD even in the ca of 2–3loci incompatibility.At our immunohematology department,a CB bank is active and clo to 1000typed CB are banked.In our department,a ction for the preparation and clinical application of cryo-platelet glue is active as well [5]and,becau of the quality of the results obtained over time,the number of requests for application in different dia condition is increas-ing.The success rate is exceeding 85%with wound healingin patients refractory to conventional
*
Correspondingauthor.Tel.:+39-10-352-822;fax:+39-10-555-6807.
E-mail address:***********************************.it,**************************.it (M.Valbonesi).
1473-0502/$-e front matter Ó2004Elvier Ltd.All rights rerved.
doi:10.ansci.2003.11.006
/journals/tras
Transfusion and Apheresis Science 30(2004)
153–156
treatment,surgery included.For the uncommon patients who fail our cryo-platelet glue it is felt that unresponsiveness is due to the lack of specific tissue stem cells or their unresponsiveness to nor-mal appropriate stimuli[6–8]or to the inability of circulatingmultipotent uncommitted stem cells to reach the place where they have to differentiate[9]. In this line is the report by Badiavas and Falanga
who successfully treated chronic wound with bone marrow-derived cells[10].To face the problem of chronic wounds unresponsive to therapy,cryo-platelet glue included,it was felt by our group that local injection of HLA compatible(2mismatches accepted)cord blood cells was an appropriate therapeutic attempt.
2.Material and methods
Two patients were enrolled after appropriate written connt after failure of adequate multiple cours of medical and/or surgical treatment.The demographic of the patients is summarized in Table1.
The autologousfibrin-platelet glue(FPG)was obtained and ud as previously described[5]. From each donation2aliquots of cryoprecipitate and4of dry platelets were obtained.After prep-aration,1aliquot of cryoprecipitate and3of platelets were frozen and kept at)80°C until ud. For the preparation of the gel,one aliquot of cryoprecipitate and one of platelets are mixed and 0.5gof calcium g luconate dissolved in3–4vials of Botropa are added.Botropa(Ravizza Farma-ceutici S.p.A,Milano,Italia)is a thrombin-like drugcommercially available that contains Bot-roxobin,a Rusll’s viper venom component,that promotes coagulation through factor X activation and platelet
activation.Thefibrin-platelet glue is prepared at the bedside in Petri dishes,cut, moulded and ud as needed.Cord blood cells collected and stored accordingto FACH T stan-dards[6]were typed employingSSP/SSO re-agents.
For class I the ABC and for class II the DR and DQ loci were typed.When needed,quencingwas carried out usingABI PRISM310Genetic Ana-lyzer(Applied Biosystems,Foster City,CA,USA). CD34+cells in the cord blood units were mea-sured usinga Facscan apparatus(Becton–Dickin-son,S.Jo,Canada)and reagents.WBC counts were obtained by a cell Coulter(ADVIA e120 Hematology system,Bayer,Tarrytown,NY, USA).Sterility was evaluated usingBACTEC e BD Peds Plus/F(Becton–Dickinson,S.Jo, Canada).
Cord blood cells stored in liquid Nitrogen were thawed a+37°C,centrifuged and the buffy-coat divided into3and4aliquots respectively for suc-cessive weekly applications.Each aliquot con-taininga minimum of26·103CD34+cells and of 12·106WBC after washingin saline was injected in the margins of the skin lesion,followed by the application of the FPG.
3.Results
Two concutive patients were included in our preliminary study,after they failed standard con-ventio
nal therapies for longer then1year.In Table 2the clinical results of treatment are reported alongwith the treatment modalities.Patient number2had also failed with repeat cryo-platelet gel application.When included in the study,the skin lesion was clearly infected and no granulation tissue was en.After the cond application of CB followed by FPG the lesion was cleared with no
Table1
Characteristics of the2patients submitted to CB
Patients Age/Gender Diagnosis Previous therapies Size(cm)of the
skin lesion Number of HLA mismatches
182/M Trauma Debridement11·71in locus C
276/F Radiation injury Local cryo-platelet-gel8·31in locus B and1in locus C Cryo-platelet glue treatment.
154M.Valbonesi et al./Transfusion and Apheresis Science30(2004)153–156
sign of infection.In this as in the other ca improvement was anticipated by incread vascu-larity of the perilesional skin and formation of granulation tissue.
No sign of graft versus tissue reactions was en in a follow-up period of3–7months.
4.Comment
Cell-bad therapies will benefit from a source of pluripotent stem cells alternative to the autol-ogous ones,who collection has potential limita-tions such as the age of the patient,his cardiac condition and the need for general anaesthesia for bone marrow aspiration.Cord blood cells are known to contain pluripotent cells which can dif-ferentiate into different cell lines in the prence of lineage specific induction factors.Cord blood cells are collected easily and are ready for clinical u when an appropriate request is made.Their responsibility in determiningg raft versus tissue reaction is modest and,given the reports of their plasticity,it is conceivable that they can produce new skin cells or rejuvenate nescentfibroblasts unresponsive to locally produced or external administration of transformingg rowth factors or cytokines.In keepingwith this is the very recent report of the successful u of bone marrow-de-rived cells in the treatment of non healingchronic wounds in3patients with wounds of more than1 year duration in whom there was histologic evi-dence sugges
ting engraftment of applied autolo-gous cells[10].The2patients in our ca list got resolution of their wounds even though patient no. 2had been included in a report from our group as the sole negative result with FPG[5].Of cour, there are limitations in our very experimental study.We have not been able,prently,to dem-onstrate that there was a change from CD34+ cells injected into skin progenitor cells.We have only en a dramatic increa in cellularity and in immature cells in the site of the cord blood injec-tions.We cannot state if a rejuvenation of nes-cent localfibroblasts took place instead of a colonization by cord stem cells condary to their plasticity.We have only shown what the feasibility of this treatment should be if it is not complicated by graft versus tissue reactions when up to2HLA mismatches are accepted between the cord blood cells and the recipient.We have also shown that, from a clinical point of view,optimal results are met even in patients unresponsive to conventional therapies and to the local application of cryo-platelet gels.On the basis of the results we are planninga larg er study after an appropriate con-nt by our local ethical committee. References
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Table2
Treatment modality and clinical results
Patient CB Applications No of CD
34+·103/appl No of WBC·
106/appl
FPG application CR(scored0–4)
13301434
24261233
CB:cord blood;FPG:cryo-platelet gel;CR:clinical result:0no improvement,1clear but modest,2partial,3almost complete,4 complete resolution of the skin lesion.
M.Valbonesi et al./Transfusion and Apheresis Science30(2004)153–156155
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