Exploring hematopoietic stem cell population in human milk and its benefits for infants:A scoping review

Objective:To comprehensively explore hematopoietic stem cells(HSCs)in human milk,understanding their molecular markers,isolation methods,benefits for infants,and potential medical applications.Methods:We conducted a scoping literature review following the PRISMA-ScR guidelines.This review included studies investigating HSCs in human milk,utilizing molecular markers such as CD34^(+),CD113^(+),and CD117^(+)for characterization.Both in vitro and in vivo studies exploring the morphology,function,and clinical implications of these cells were considered.The diverse range of papers reviewed were indexed in PubMed,Science Direct,Scopus,Sage Journals,and Google Scholar,published between 2010 and 2023.Results:This scoping review explored 577 articles and selected 13 studies based on our inclusion criteria,focusing on HSCs in human milk.Most studies dilute samples prior to HSC isolation,followed by detection using markers such as CD34^(+),CD113^(+),and CD117^(+),with flow cytometry serving as the primary analysis tool,focusing on their isolation and detection methods.While no definitive benefits have been conclusively established,there is a strong belief in the potential of HSCs to positively impact infant immunity,growth,and tissue repair.Conclusions:This review presents significant evidence supporting the presence of HSCs in human milk,identified by markers such as CD34^(+),CD113^(+),and CD117^(+).These cells show considerable potential in enhancing infant health,including immunity,tissue repair,cognitive development,and gastrointestinal health.Despite methodological variations in isolation and detection techniques,the collective findings underscore the potential clinical relevance of HSCs in human milk.Moreover,this review highlights the noninvasive accessibility of human milk as a source of HSCs and emphasizes the need for further research to unlock their therapeutic potential.

Hematopoietic stem cells in research and clinical applications:The“CD34 issue”

In this paper,experimental findings concerning the kinetics of hematopoietic reconstitution are compared to corresponding clinical data.Although not clearly apparent,the transplantation practice seems to confirm the basic proposals of experimental hematology concerning hematopoietic reconstitution resulting from successive waves of repopulation stemming from different subpopulations of progenitor and stem cells.One of the "f irst rate" parameters in clinical transplantations in hematology;i.e.the CD34+ positive cell dose,has been discussed with respect to the functional heterogeneity and variability of cell populations endowed by expression of CD34.This parameter is useful only if the relative proportion of stem and progenitor cells in the CD34+ cell population is more or less maintained in a series of patients or donors.This proportion could vary with respect to the source,pathology,treatment,processing procedure,the graft ex vivo treatment and so on.Therefore,a universal dose of CD34+ cells cannot be def ined.In addition,to avoid further confusion,the CD34+ cells should not be named "stem cells" or "progenitor cells" since these denominations only concern functionally characterized cell entities.

HUMAN PRIMITIVE HEMATOPOIETIC PROGENITOR CELLS ARE MORE ENRICHED IN CD34^+CD38^- POPULATION THAN IN CD34^+CD38^+ POPULATION

To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fractions in CD34+cells with or without CD38 expression. A clonogenic assay showed that most CFC were present in CD34+CD38+ population. Morphologic analysis showed that blast-like cells were more enriched in the CD34+CD38 fraction. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell function by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At the first two weeks, more CF.C harvested from the culture in the fractions initiated with both populations. However, more LTC-IC were present during weeks 4 to 12 in the CD34+CD38- population. These results indicate the primitive progenitors are more enriched in CD34+CD38 population than in CD34+CD38+ cells.

Autologous CD34^+ and CD133^+ stem cells transplantation in patients with end stage liver disease

AIM:To assess the utility of an autologous CD34 + and CD133 + stem cells infusion as a possible therapeutic modality in patients with end-stage liver diseases.METHODS:One hundred and forty patients with endstage liver diseases were randomized into two groups.Group 1,comprising 90 patients,received granulocyte colony stimulating factor for five days followed by autologous CD34 + and CD133 + stem cell infusion in the portal vein.Group 2,comprising 50 patients,received regular liver treatment only and served as a control group.RESULTS:Near normalization of liver enzymes and improvement in synthetic function were observed in 54.5% of the group 1 patients;13.6% of the patients showed stable states in the infused group.None of the patients in the control group showed improvement.No adverse effects were noted.CONCLUSION:Our data showed that a CD34 + and CD133 + stem cells infusion can be used as supportive treatment for end-stage liver disease with satisfactory tolerability.

Donor-Derived CD19-Targeted T Cell Infusion Eliminates B Cell Acute Lymphoblastic Leukemia Minimal Residual Disease with No Response to Donor Lymphocytes after Allogeneic Hematopoietic Stem Cell Transplantation

Leukemia relapse is still the leading cause of treatment failure after allogeneic hematopoietic stem cell transplantation (allo-HSCT) for B cell acute lymphoblastic leukemia (B-ALL). Relapsed patients with BALL after allo-HSCT have a very short median survival. Minimal residual disease (MRD) is predictive of forthcoming hematological relapse after hematopoietic stem cell transplantation (HSCT);furthermore, eliminating MRD effectively prevents relapse. Donor lymphoblastic infusion (DLI) is the main established approach to treat B-ALL with MRD after allo-HSCT. However, about one-third of patients with MRD are non-responsive to DLI and their prognosis worsens. Although donor-derived cluster of differentiation (CD)19-directed chimeric antigen receptor-modified (CAR) T cells (CART19s) can potentially cure leukemia, the efficiency and safety of infusions with these cells have not yet been investigated in patients with MRD after HSCT. Between September 2014 and February 2018, six patients each received one or more infusions of CART19s from HSCT donors. Five (83.33%) achieved MRD-negative remission, and one case was not responsive to the administration of CAR T cells. Three of the six patients are currently alive without leukemia. No patient developed acute graft-versus-host disease (aGVHD), and no patient died of cytokine release syndrome. Donor-derived CAR T cell infusions seem to be an effective and safe intervention for patients with MRD in B-ALL after allo-HSCT and for those who were not responsive to DLI.

Impact of T cells on hematopoietic stem and progenitor cell function:Good guys or bad guys?

When hematopoietic stem and progenitor cells(HSPC)are harvested for transplantation, either from the bone marrow or from mobilized blood, the graft contains a significant number of T cells. It is these T cells that are the major drivers of graft-vs-host disease(Gv HD). The risk for Gv HD can simply be reduced by the removal of these T cells from the graft. However, this is not always desirable, as this procedure also decreases the engraftment of the transplanted HSPCs and, if applicable, a graft-vs-tumor effect. This poses an important conundrum in the field: T cells act as a double-edged sword upon allogeneic HSPC transplantation, as they support engraftment of HSPCs and provide anti-tumor activity, but can also cause Gv HD. It has recently been suggested that T cells also enhance the engraftment of autologous HSPCs, thus supporting the notion that T cells and HSPCs have an important functional interaction that is highly beneficial, in particular during transplantation. The underlying reason on why and how T cells contribute to HSPC engraftment is still poorly understood. Therefore, we evaluate in this review the studies that have examined the role of T cells during HSPC transplantation and the possible mechanisms involved in their supporting function. Understanding the underlying cellular and molecular mechanisms can provide new insight into improving HSPC engraftment and thus lower the number of HSPCs required during transplantation. Moreover, it could provide new avenues to limit the development of severe Gv HD, thus making HSPC transplantations more efficient and ultimately safer.

<i>In vitro</i>differentiation of human umbilical cord-derived mesenchymal stem cells into CD34⁺cells via CD34 antibody

CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentiation from MSCs, especially from human MSCs (hUC-MSCs) are still unclear. In the current study, the effect of CD34 antibody on the CD34+ cell differentiation from human umbilical cord (UC)-derived MSCs (hUC-MSCs) is determined. The results have demonstrated that the expression of cd34 protein is significantly increased in hUC-MSCs treated with CD34 antibody. In addition, the cell proliferation is increased in hUC-MSCs after treatment with CD34 antibody. Moreover, the expression of PI3K, AKT, p-AKT proteins, which are signaling molecules related to stem cell differentiation, is increased by CD34 antibody. The results suggest that CD34 antibody could promote the differentiation of hUC-MSCs into CD34+ cells and PI3K/AKT may be involved in this important process.

High frequency of CD34+CD38-/low immature leukemia cells is correlated with unfavorable prognosis in acute myeloid leukemia

AIM To evaluate the importance of the CD34+CD38-cell population when compared to the CD34+CD38+/low and CD34+CD38+/high leukemic cell sub-populations and to determine its correlations with leukemia characteristics and known prognostic factors, as well as with response to therapy and survival.METHODS Two hundred bone marrow samples were obtained at diagnosis from 200 consecutive patients with newly diagnosed acute myeloid leukemia(AML) were studied between September 2008 and December 2010 at our Institution(Hematology Department, Lyon, France). The CD34/CD38 cell profile was analyzed by multiparameter flowcytometry approach using 8 C panels and FACS CANTO and Diva software(BD Bioscience).RESULTS We analyzed CD34 and CD38 expression in bone marrow samples of 200 AML patients at diagnosis, and investigated the prognostic value of the most immature CD34+CD38-population. Using a cut-off value of 1% of CD34+CD38-from total "bulk leukemic cells" we found that a high(> 1%) level of CD34+CD38-blasts at diagnosis was correlated with advanced age, adverse cytogenetics as well as with a lower rate of complete response after induction and shorter disease-free survival. In a multivariate analysis considering age, leukocytosis, the % of CD34+ blasts cells and the standardized cytogenetic and molecular risk subgroups, a percentage of CD34+CD38-leukemic cells > 1% was an independent predictor of DFS [HR = 2.8(1.02-7.73), P = 0.04] and OS [HR = 2.65(1.09-6.43), P = 0.03].CONCLUSION Taken together, these results show that a CD34/CD38 "backbone" for leukemic cell analysis by multicolour flowcytometry at diagnosis provides useful prognostic information.

Human umbilical cord mesenchymal stem cells as treatment of adjuvant rheumatoid arthritis in a rat model

AIM:To investigate the effect of human umbilical cord stem cells,both mesenchymal and hematopoietic(CD34+),in the treatment of arthritis.METHODS:Mesenchymal stem cells(MSCs) and hematopoietic(CD34+) stem cells(HSC) were isolated from human umbilical cord blood obtained from the umbilical cord of healthy pregnant donors undergoing fullterm normal vaginal delivery.MSC,HSC,methotrexate(MTX) and sterile saline were injected intra-articularly into the rat hindpaw with complete freunds adjuvant(CFA) induced arthritis after the onset of disease(day 34),when arthritis had become well established(arthritis score ≥ 2).Arthritic indices were evaluated and the levels of interleukin(IL)-1,tumor necrosis factor(TNF)-α and interferon(IFN)-γ and anti-inflammatory cytokine IL-10 in serum were determined using enzyme-linked immunosorbent assay.Animals of all groups were sacrificed 34 d after beginning treatment,except positive control(PC) which was sacrificed at 10,21 and 34 d for microscopic observation of disease progression.We used hematoxylin,eosin and Masson's trichrome stains for histopathological examination of cartilage and synovium.RESULTS:The mean arthritis scores were similar in all groups at 12 and 34 d post immunization,with no statistical significant difference.Upon the injection of stem cells(hematopoietic and mesenchymal),the overall arthritis signs were significantly improved around 21 d after receiving the injection and totally disappeared at day 34 post treatment in MSC group.Mean hindpaw diameter(mm) in the MSC rats was about half that of the PC and MTX groups(P = 0.007 and P = 0.021,respectively) and 0.6 mm less than the HSC group(P = 0.047),as indicated by paw swelling.Associated with these findings,serum levels of TNF-α,IFN-γ and IL-1 decreased significantly in HSC and MSC groups compared to PC and MTX groups(P < 0.05),while the expression of IL-10 was increased.Histopathological examination with H and E stain revealed that the MTX treated group showed significant reduction of leucocytic infiltrate and hypertrophy of the synovial tissue with moderate obliteration of the joint cavity.Stem cells treated groups(both hematopoietic CD34+ and mesenchymal),showed significant reduction in leucocytic infiltrate and hypertrophy of the synovial tissue with mild obliteration of the joint cavity.With Masson's trichrome,stain sections from the PC group showed evidence of vascular edema of almost all vessels within the synovium in nearly all arthritic rats.Vacuoles were also visible in the outer vessel wall.The vessel became hemorrhagic and finally necrotic.In addition,there was extensive fibrosis completely obliterating the joint cavity.The mean color area percentage of collagen in this group was 0.324 ± 0.096,which was significantly increased when compared to the negative control group.The mean color area percentage of collagen in hematopoietic CD34+ and mesenchymal groups was 0.176 ± 0.0137 and 0.174 ± 0.0197 respectively,which showed a marked decrement compared to the PC group,denoting a mild increase in synovial tissue collagen fibers.CONCLUSION:MSC enhance the efficacy of CFAinduced arthritis treatment,most likely through the modulation of the expression of cytokines and amelioration of pathological changes in joints.

Adipose-derived mesenchymal stromal/stem cells: An update on their phenotype in vivo and in vitro

Adipose tissue is a rich, ubiquitous and easily acces-sible source for multipotent stromal/stem cells and has, therefore, several advantages compared to other sourc-es of mesenchymal stromal/stem cells. Several studies have tried to identify the origin of the stromal/stem cell population within adipose tissue in situ. This is a complicated attempt because no marker has currently been described which unambiguously identifies native adipose-derived stromal/stem cells(ASCs). Isolated and cultured ASCs are a non-uniform preparation consisting of several subsets of stem and precursor cells. Cultured ASCs are characterized by their expression of a panel of markers(and the absence of others), whereas their in vitro phenotype is dynamic. Some markers were ex-pressed de novo during culture, the expression of some markers is lost. For a long time, CD34 expression was solely used to characterize haematopoietic stem and progenitor cells, but now it has become evident that it is also a potential marker to identify an ASC subpopula-tion in situ and after a short culture time. Nevertheless, long-term cultured ASCs do not express CD34, perhaps due to the artificial environment. This review gives an update of the recently published data on the origin and phenotype of ASCs both in vivo and in vitro. In addition, the composition of ASCs(or their subpopula-tions) seems to vary between different laboratories andpreparations. This heterogeneity of ASC preparationsmay result from different reasons. One of the main problems in comparing results from different laborato-ries is the lack of a standardized isolation and culture protocol for ASCs. Since many aspects of ASCs, suchas the differential potential or the current use in clinical trials, are fully described in other recent reviews, this review further updates the more basic research issues concerning ASCs' subpopulations, heterogeneity andculture standardization.

Autologous mobilized peripheral blood CD34^+ cell infusion in non-viral decompensated liver cirrhosis

AIM: To study the effect of mobilized peripheral blood autologous CD34 positive(CD34+) cell infusion in patients with non-viral decompensated cirrhosis.METHODS: Cirrhotic patients of non-viral etiology were divided into 2 groups based on their willingness to be listed for deceased donor liver transplant(DDLT)(control, n = 23) or to receive autologous CD34+ cell infusion through the hepatic artery(study group, n= 22). Patients in the study group were admitted to hospital and received granulocyte colony stimulating factor injections 520 μg/d for 3 consecutive days to mobilize CD34+ cells from the bone marrow. On day 4,leukapheresis was done and CD34+ cells were isolated using CliniMAC magnetic cell sorter. The isolated CD34+ cells were infused into the hepatic artery under radiological guidance. The patients were discharged within 48 h. The control group received standard of care treatment for liver cirrhosis and were worked up for DDLT as per protocol of the institute. Both groups were followed up every week for 4 wk and then every month for 3 mo.RESULTS: In the control and the study group, the cause of cirrhosis was cryptogenic in 18(78.2%) and16(72.72%) and alcohol related in 5(21.7%) and6(27.27%), respectively. The mean day 3 cell count(cells/μL) was 27.00 ± 20.43 with a viability of 81.84± 11.99%. and purity of 80%-90%. Primary end point analysis revealed that at 4 wk, the mean serum albumin in the study group increased significantly(2.83± 0.36 vs 2.43 ± 0.42, P = 0.001) when compared with controls. This improvement in albumin was,however, not sustained at 3 mo. However, at the end of3 mo there was a statistically significant improvement in serum creatinine in the study group(0.96 ± 0.33 vs 1.42 ± 0.70, P = 0.01) which translated into a significant improvement in the Model for End-Stage Liver Disease score(15.75 ± 5.13 vs 19.94 ± 6.68,P = 0.04). On statistical analysis of secondary end points, the transplant free survival at the end of 1 mo and 3 mo did not show any significant difference(P =0.60) when compared to the control group. There was no improvement in aspartate transaminase, alanine transaminase, and bilirubin at any point in the study population. There was no mortality benefit in the study group. The procedure was safe with no procedural or treatment related complications.CONCLUSION: Autologous CD 34+ cell infusion is safe and effectively improves liver function in the short term and may serve as a bridge to liver transplantation.

Radix Ilicis Pubescentis total flavonoids combined with mobilization of bone marrow stem cells to protect against cerebral ischemia/reperfusion injury

Previous studies have shown that Radix Ilicis Pubescentis total flavonoids have a neuroprotective effect, but it remains unclear whether Radix Ilicis Pubescentis total flavonoids have a synergistic effect with the recombinant human granulocyte colony stimulating factor-mobilized bone marrow stem cell transplantation on cerebral ischemia/reperfusion injury. Rat ischemia models were administered 0.3, 0.15 and 0.075 g/kg Radix Ilicis Pubescentis total flavonoids from 3 days before modeling to 2 days after injury. Results showed that Radix Ilicis Pubescentis total flavonoids could reduce pathological injury in rats with cerebral ischemia/reperfusion injury. The number of Nissl bodies increased, Bax protein expression decreased, Bcl-2 protein expression increased and the number of CD34-positive cells increased. Therefore, Radix Ilicis Pubescentis total flavonoids can improve the bone marrow stem cell mobilization effect, enhance the anti-apoptotic ability of nerve cells, and have a neuroprotective effect on cerebral ischemia/reperfusion injury in rats.

Establishment and molecular characterization of human dermal mesenchymal-like stem cells derived from human scalp biopsy of androgenetic alopecia patient

Development of Dermal cell line has great scope in the field of skin related diseases and regenerative medicine. Alopecia leads to a skin disorder causing balding and its mechanism is not yet understood. In the present study, we have developed and characterized a heterogeneous population of human dermal mesenchymal-like stem cell line from scalp biopsy of androgenetic alopecia patient with a view to isolate cells from the bulge region of the hair follicle. Our study showed that the dermal cells isolated from dermis of skin showed epithelial-like cells expressing CD34 and Keratin 18, which are characteristic of bulge hair follicle cells. These cells also expressed mesenchymal phenotypes and pluripotency markers such as Oct4, Nanog and SOX2. These cells were designated as “Human Dermal Mesenchymal-like Stem Cells (hDMSCs)”. To confirm their epithelial phenotypes, we have grown these cells at low serum concentration and it was observed that 3% serum concentration provided optimum conditions for their growth and maintenance of characteristics. The hDMSCs cells are presently at passage 10. This study reports the establishment of human dermal mesenchymal-like cell line from the dermis of Alopecia patient, which may be used as an in vitro model system to study the mechanism of Alopecia and other related skin disorders.

Identification of Bulge Stem Cells in Mouse and Human Hair Follicles

The skin contains various populaions of stem cells, but its characterization has been hampered by lack of markers and unclear location. The hair follicle has a niche for stem cells called a “bulge” which acts as a reservoir of multipotent stem cells. In the study reported here, an immunohistochemical and immunofluorescence analysis was performed on mouse and human tissues in order to determine the possible presence of stem cells of hair follicle through cytokeratin 15 (CK15), CD34, and CD200 markers identified as crucial to the stem cells and to identify the bulge region. Mouse (n = 7) and human (n = 7) skin samples were used. The expression of proteins was determined by the indirect immunoperoxidase technique and a secondary antibody bound to a fluorochrome. The specificity of staining was evaluated by negative controls. The results revealed that the stem cells associated with CD34 and CD200 antibodies were differentially expressed in the interfollicular epidermis, sebaceous glands, and bulge region, indicating that, in mice, CD34 and, in humans, CD200 are more specific than CK15 in detecting bulge cells. It also suggests that CD34 is specific for mouse bulge cells, while CD200 might have specificity for progenitor cells and partially differentiated cells in humans.

Autologous Peripheral Blood Stem Cells and <i>γ</i>/<i>δ</i>T Cells May Improve Immunity in Treating Secondary Bacteremic Infection in HIV Infected Patient

Opportunistic bacteremia in adult HIV-infected patients is a normal co-infectious condition caused by Gram-negative bacilli. Respiratory infections, including cough, shortness of breath, and chest pain and skin infection with eruptions, pustules and itchiness, are common complaints in the setting of late HIV infection. The variety of infections ranges from mild, self-limited viral, bacteremia and fungal infections to severe, life-threatening demanding urgent care and hospitalization. Varicella pneumonia, for instance, is the most severe complication of chickenpox in HIV infected adults, potentially refractory, fulminant respiratory failure can ensue. Patients with impaired immune status and chronic lung disease are at an increased risk. In the United States as well as in Vietnam, bacterial/viral pneumonia and skin infection are the two most common HIV-associated conditions. While globally the incidence of opportunistic infection has decreased since the introduction of highly active antiretroviral therapy during the last 3 decades, HIV-associated diseases remain a significant source of mortality, thus any manifestation must be taken seriously. This study will present the most common HIV-related pulmonary and skin infections and provide an overview of the epidemiology, characteristic clinical and chest radiograph findings, diagnosis, treatment, and prevention globally as well in Vietnam. Though the extensive efforts of the Vietnamese Government during last decade contributed to a valuable decrease, yet epidemic in Vietnam still remains high, ranking Vietnam 5th in the South-East region. The second part of the study focuses on a unique and severe HIV case report of a 35-year-old man, with a rare form of pneumonia caused by Acitenobacter spp. concomitant with a prolonged and disseminating skin infection. The case has been treated with a combination of conventional anti-retroviral medication and autologous peripheral blood stem cells, the results showed that within 5 months there was an impressive amelioration of HIV viral activity together with a total recovery from pneumonia and skin infection.

应用免疫磁珠法分离脐血CD_(34)^+造血细胞

目的：分离脐血CD细胞，以了解其造血增殖活性。方法：应用免疫磁珠法（Isolex50系统）分离脐血CD细胞，并对其实验方法进行了改进。结果：常规方法CD纯度较低，经过改进，脐血CD细胞由分离前的（1．39±0．76）％增加至（59．46±17．35）％，CD细胞富集倍数为（62．31±55．19）倍。分离后组分CFU－GM、BFU－E、CFU－Mix较分离前分别增加了（41．05±15．77）、（13．14±8.66）及（34．80±19．58）倍，而阴性组分所形成的集落显著减少。结论：免疫磁珠法可以有效地分离CD细胞，造血祖细胞集落大多来源于CD细胞。

肿瘤患者外周血干细胞动员过程中CD_(34)^+细胞的动态变化及意义

为观察肿瘤患者 CD+ 34 细胞数量在动员过程中的动态变化 ,确定采集干 /祖细胞的最佳时机 ,采用环磷酰胺 (CTX)联合粒细胞集落刺激因子 (G- CSF)对 2 1例肿瘤患者进行造血干 /祖细胞动员。动员过程中 ,每天进行血细胞计数 ,于前期、极期、恢复早期、恢复期计数外周血 CD+ 34 细胞、白细胞 (WBC)、单核细胞 (MNC)、血小板(Plt)。结果显示 ,不同患者出现极期、恢复早期、恢复期的时间差异较大。与前期相比 ,极期 CD+ 34 明显降低(P<0 .0 5 ) ,恢复期则显著增加 (P<0 .0 1)。WBC、MNC、Plt变化均与 CD+ 34 细胞变化呈显著正相关 (r分别为 0 .99、0 .82 7、0 .886 ,P均 <0 .0 1)。提示在 WBC>5 .0× 10 9/ L时采集外周血造血干细胞 (PBSC)较合适 ,MNC>1.5×10 9/ L时采集 PBSC较理想 ,监测 Plt计数变化 。

CD_(34)定量分析外周血干细胞的研究

目的 定量检测外周血干细胞 (PBSC)。方法 以藻红蛋白 (PE)标记的CD34单克隆抗体 ,用流式细胞术 (FCM)对 34例白血病患者与 33例健康成人外周血表达CD34抗原 (CD34+)的细胞进行定量分析 ,并以普通光学显微镜 (LM )、透射电子显微镜 (TEM)、激光扫描共聚焦显微镜 (LSCM )分别对CD34+/CD34-细胞进行观察。结果 经LSCM证实 ,FCM定量检测CD34+细胞特异性强 ,CD34+细胞在 0 %～ 1 6 .1 %范围内线性良好 (r =0 .9947) ,当CD34+细胞为 91 0 %、38 3%时 ,CV <3%。CD34+/CD34-细胞在LM、TEM下形态与结构相似 ,但CD34+细胞在LSCM下显示圆环状橙红色荧光。结论 FCM检测外周血干细胞灵敏、特异、快速 。

人造血干/祖细胞多态性的研究:Ⅷ.人正常骨髓 CD34^+造血细胞体视学的特征

人CD34^+造血细胞是具有高度自我更新、多向分化及重建长期造血与免疫学功能的独特体细胞。为系统探索CD34^+造血细胞的形态、细胞化学及超微结构特征,新近我们设计组合并建立了CIMS-100-FACS 440无菌二次分选术,可使所获CD34^+造血细胞的纯度达100%。在此基础上,本研究采用Cambri-dge Quantimet 970全自动图像分析仪对光学显微镜、扫描电镜及透射电镜下的CD34^+造血细胞进行了体视学方面的某些探讨,进一步从三维结构信息中深刻揭示CD34^+造血细胞的形态计量学特征。经扫描→模数转换←阴影校正→图像暂存←统计分析等检测,结果表明:CD34^+造血细胞的直径3.490—6.741μm,周长11.776—26.240μm,面积9.565—35.686μm^2,形状因子1.048—1.840,核浆比0.58—0.72,平均光密度0.17675—0.65100,积分光密度2717.217—9870.643。由此可见CD34^+造血细胞的确为非均一细胞群,这可能与CD34^+造血细胞的功能亚群与分化阶段密切相关。据我们所知,这是国际上首次有关人CD34^+造血细胞体视学特征的报道。