scholarly journals Flow cytometric analysis of normal human megakaryocytes

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1244-1252 ◽  
Author(s):  
A Tomer ◽  
LA Harker ◽  
SA Burstein
Keyword(s):  
Flow Cytometry ◽  
Cell Membrane ◽  
Cell Size ◽  
Cell Structure ◽  
Surface Membrane ◽  
Flow Cytometric Analysis ◽  
Light Scatter ◽  
Membrane Expression ◽  
Color Flow ◽  
The Mean

Megakaryocytes from normal routine human bone marrow aspirates were analyzed by flow cytometry for size, fine cell structure and granularity, membrane expression of glycoprotein (GP) IIb/IIIa and ploidy. Marrow cells were initially enriched for megakaryocytes by a Percoll density gradient and megakaryocytes were labeled with a fluoresceinated monoclonal antibody directed to the GPIIb/IIIa complex. The cells were fixed with paraformaldehyde and stained with propidium iodide (PI) for DNA quantitation. Using two-color flow cytometry, megakaryocytes were identified by their high membrane immunofluorescence and their ploidy was determined according to the relative fluorescence intensity of the PI. Forward light scatter (FSC), correlating with cell size, 90 degrees side light scatter (SSC), reflecting primarily cell internal fine structure and granularity, and total cell membrane fluorescence were examined. To evaluate independently the relationship between size and cell membrane fluorescence obtained by flow cytometry, megakaryocytes were sorted directly on slides and analyzed by a laser-based anchored cell analyzer (ACAS). There was a strong correlation among size, SSC, and the level of membrane fluorescence. The mean diameter of megakaryocytes was 28.1 +/- 12.3 micron. The modal ploidy distribution was 16N with approximately one-fifth of the cells less than or equal to 4N. The mean FSC and SSC levels increased with increasing ploidy. However, the marked overlap observed between the ranges of these parameters in adjacent ploidy classes suggested that size and SSC increase continuously rather than by discrete steps as is characteristic for ploidy. The total surface membrane fluorescence was correlated with cell size (r = 0.98) as measured by FSC or directly by the ACAS (r = 0.85), and with cell ploidy (r = 0.99) indicating an augmentation in total membrane GPIIb/IIIa expression with an increase in cell size and ploidy. However, estimated GPIIb/IIIa fluorescence density was inversely correlated with FSC suggesting that the GPIIb/IIIa surface epitope density is decreased with increasing cell maturity. We conclude that flow cytometry is a useful technique for the rapid analysis of human megakaryocytes obtained by marrow aspiration, and should be applicable to studies of pathologic states.

scholarly journals Flow cytometric analysis of normal human megakaryocytes

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1244-1252 ◽  
Author(s):  
A Tomer ◽  
LA Harker ◽  
SA Burstein
Keyword(s):  
Flow Cytometry ◽  
Cell Membrane ◽  
Cell Size ◽  
Cell Structure ◽  
Surface Membrane ◽  
Flow Cytometric Analysis ◽  
Light Scatter ◽  
Membrane Expression ◽  
Color Flow ◽  
The Mean

Abstract Megakaryocytes from normal routine human bone marrow aspirates were analyzed by flow cytometry for size, fine cell structure and granularity, membrane expression of glycoprotein (GP) IIb/IIIa and ploidy. Marrow cells were initially enriched for megakaryocytes by a Percoll density gradient and megakaryocytes were labeled with a fluoresceinated monoclonal antibody directed to the GPIIb/IIIa complex. The cells were fixed with paraformaldehyde and stained with propidium iodide (PI) for DNA quantitation. Using two-color flow cytometry, megakaryocytes were identified by their high membrane immunofluorescence and their ploidy was determined according to the relative fluorescence intensity of the PI. Forward light scatter (FSC), correlating with cell size, 90 degrees side light scatter (SSC), reflecting primarily cell internal fine structure and granularity, and total cell membrane fluorescence were examined. To evaluate independently the relationship between size and cell membrane fluorescence obtained by flow cytometry, megakaryocytes were sorted directly on slides and analyzed by a laser-based anchored cell analyzer (ACAS). There was a strong correlation among size, SSC, and the level of membrane fluorescence. The mean diameter of megakaryocytes was 28.1 +/- 12.3 micron. The modal ploidy distribution was 16N with approximately one-fifth of the cells less than or equal to 4N. The mean FSC and SSC levels increased with increasing ploidy. However, the marked overlap observed between the ranges of these parameters in adjacent ploidy classes suggested that size and SSC increase continuously rather than by discrete steps as is characteristic for ploidy. The total surface membrane fluorescence was correlated with cell size (r = 0.98) as measured by FSC or directly by the ACAS (r = 0.85), and with cell ploidy (r = 0.99) indicating an augmentation in total membrane GPIIb/IIIa expression with an increase in cell size and ploidy. However, estimated GPIIb/IIIa fluorescence density was inversely correlated with FSC suggesting that the GPIIb/IIIa surface epitope density is decreased with increasing cell maturity. We conclude that flow cytometry is a useful technique for the rapid analysis of human megakaryocytes obtained by marrow aspiration, and should be applicable to studies of pathologic states.

scholarly journals Comparative analysis of surface membrane immunoglobulin determination by flow cytometry and fluorescence microscopy.

1986 ◽  
Vol 34 (4) ◽  
pp. 475-481 ◽  
Author(s):  
D F Gebhard ◽  
A Mittelman ◽  
C Cirrincione ◽  
H T Thaler ◽  
B Koziner
Keyword(s):  
Flow Cytometry ◽  
Standard Deviation ◽  
Fluorescence Microscopy ◽  
B Cell ◽  
Membrane Surface ◽  
Surface Membrane ◽  
Flow Cytometric Analysis ◽  
Normal Individuals ◽  
The Mean ◽  
Hodgkin's Lymphomas

The analysis of membrane surface immunoglobulin (SmIg) on B lymphocytes was carried out in 59 normal individuals and nine patients with B-cell non-Hodgkin's lymphomas by conventional immunofluorescence microscopy and flow cytometry. Five channel settings of a cytofluorograph were evaluated (100, 150, 200, 250, 300) and the mean and standard deviation of the percent positive cells were calculated and compared to the mean and standard deviation of the microscope reading. On the basis of the relative fluorescence reactivity, we were able to determine a fluorescence intensity at which the results of flow cytometry and fluorescence microscopy were comparable. In normal individuals, for cells expressing surface Ia, the channel giving similar results to that of fluorescence microscopy was 150; for kappa and lambda chains, channel 200; for Fab'PV, channel 200; and for IgM, channel 250. In patients with B-cell non-Hodgkin's lymphomas, for cells expressing surface Ia the channel giving similar results to that of fluorescence microscopy was 100; for kappa, channel 100; for lambda, channel 200; for Fab'FV, channel 150; and for IgM, channel 150. Flow cytometric analysis of SmIg appears to be superior to fluorescence microscopy in efficiency, and has the added advantages of being a rapid, sensitive, and objectively quantitative methodology.

Detecting Cryptic Neoplastic B-Cells: Flow Cytometric Analysis of B-Cell Lymphoma Concealed in Polyclonal B-Cells and Other Intense Reactive Components, with Emphasis on Large B-Cell Lymphoma in Fine Needle Aspirate (FNA)/ Body Fluid (BF).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4560-4560
Author(s):  
Endi Wang ◽  
Joan E. Etzell
Keyword(s):  
Flow Cytometry ◽  
B Cells ◽  
B Cell ◽  
Cell Lymphoma ◽  
Flow Cytometric Analysis ◽  
B Cell Lymphoma ◽  
Light Scatter ◽  
Color Flow ◽  
Large B Cell Lymphoma ◽  
Large B Cell

Abstract Flow cytometry is widely used in the diagnosis of lymphoma. In B-cell lymphoma, light chain restriction (LCR) is especially useful for distinguishing B-cell malignancy from a reactive lymphoid proliferation. Usually the neoplastic B-cell population predominates, and monoclonality is conspicuous as demonstrated by distinct LCR. However, if the specimen contains a large reactive component, especially polyclonal B-cells, a small number of neoplastic B-cells will be buried in the background, thus creating a diagnostic challenge. We retrospectively analyzed three/four-color flow cytometry by examining 12 cases of B-cell lymphoma with a small proportion of neoplastic B-cells hidden in polyclonal B-cells and other intense reactive components. These 12 cases (9 FNA, 1 BF, 2 tissues) comprised 7% of specimens diagnostic or suspicious for lymphoma by flow cytometry at our institution. Eight cases were diffuse large B-cell lymphoma (DLBCL) (18.6% of DLBCL and 27.6% of DLBCL in FNA/BF) while four were lymphoma of other types (3% of non-DLBCL). In all the cases, LCR was obscured by polyclonal B-cells (8/12) or absence of surface immunoglobulin (sIg) in neoplastic B-cells (4/12), and morphology resembled a reactive picture. The clues by flow cytometry prompting further analysis included increased forward angle light scatter(FSC)/side angle light scatter(SSC) (12/12), brighter CD20 (6/12), dim CD20 (1/12), absence of sIg (4/12), and a distinct population of B-cells co-expressing CD10 (2/12), CD5 (1/12), CD2 (1/12), CD23 (4/12) or CD38 (1/12). By backgating/regating suspected subpopulations, the concealed neoplastic B-cells were demonstrated by enriched LCR (5 to10 fold) or tight clustering on FSC/SSC. In conclusion, cryptic neoplastic B-cells, especially diffuse large B-cell lymphoma in FNA/BF, can be extracted, according to their altered immunophenotypic features, from background polyclonal B-cells and reactive T-cells via manipulation of gating strategies.

scholarly journals Flow cytometric analysis of megakaryocytes from patients with abnormal platelet counts

Blood ◽  
1989 ◽  
Vol 74 (2) ◽  
pp. 594-601 ◽  
Author(s):  
A Tomer ◽  
P Friese ◽  
R Conklin ◽  
W Bales ◽  
L Archer ◽  
...  
Keyword(s):  
Cell Size ◽  
Flow Cytometric Analysis ◽  
Distribution Analysis ◽  
Membrane Expression ◽  
Platelet Destruction ◽  
Platelet Production ◽  
Flow Cytometric ◽  
High Ploidy ◽  
The Right ◽  
Primary Thrombocytosis

Abstract Megakaryocytes (MKs) from 40 patients with quantitative platelet disorders and 19 normal volunteers were analyzed by flow cytometry for size, fine cell internal structure and granularity, membrane expression of the glycoprotein (GP) IIb/IIIa complex, and for ploidy distribution. Analysis was performed on unfractionated minimally manipulated marrows obtained from routine bone marrow aspirates. MKs were labeled with a fluorescent lineage-specific monoclonal antibody to the GPIIb/IIIa complex followed by DNA staining with propidium iodide. Eight hundred to 3,000 MKs were analyzed in each sample. The modal ploidy distribution in normals was 16N, comprising about half of the megakaryocytic population, with 22.6% of the cells less than or equal to 8N and 22.0% greater than or equal to 32N. Twelve thrombocytopenic patients with decreased marrow MKs on biopsy (mean platelet count [MPC] 44,600/microliters) showed an increase in low ploidy cells with 53.2% less than or equal to 8N (P less than .01); cell size was reduced in three patients when compared to normal cells of identical ploidy (P less than .05). Eight thrombocytopenic patients with enhanced platelet destruction (with normal or increased MKs on biopsy and shortened platelet survival; MPC 41,400/microliters) showed an increased proportion of high ploidy cells greater than or equal to 32N to 39.2% (P less than .01). Increased cell size and granularity were found in four of these patients (P less than .05). Six patients with thrombocytopenia secondary to multiple mechanisms affecting both platelet production and destruction (MPC 66,700/microliters) showed no shift in ploidy. Four patients with primary thrombocytosis (two with thrombocythemia and two with polycythemia vera; MPC 822,500/microliters) showed a marked shift toward high ploidy cells with 42.3% greater than or equal to 32N and 7.6% greater than or equal to 64N cells (P less than .01). The shift was accompanied by a marked increase in cell size and granularity in the patients with thrombocythemia. Ten patients with thrombocytosis secondary to chronic blood loss, malignant or inflammatory disorders (MPC 714,000/microliters), showed variable distributions with four patients exhibiting a shift in ploidy to the right similar to that found in the patients with increased platelet destruction. Based upon the present data, flow cytometric ploidy distribution may be diagnostically useful in thrombocytopenic patients by discriminating between disorders of platelet production and destruction.

Cell structure and mechanical properties of microcellular PLA foams prepared via autoclave constrained foaming

2020 ◽  
pp. 026248932093032
Author(s):  
Jinwei Chen ◽  
Ling Yang ◽  
Dahua Chen ◽  
Qunshan Mai ◽  
Meigui Wang ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Cell Size ◽  
Cell Structure ◽  
Saturation Pressure ◽  
Tensile Modulus ◽  
Cell Structures ◽  
Wide Range ◽  
The Mean ◽  
Constrained Foaming ◽  
The Relationship

Microcellular polylactic acid (PLA) foams with various cell size and cell morphologies were prepared using supercritical carbon dioxide (sc-CO2) solid-state foaming to investigate the relationship between the cell structure and mechanical properties. Constrained foaming was used and a wide range of cell structures with a constant porosity of ∼75% by tuning saturation pressure (8–24 MPa) was developed. Experiments varying the saturation pressure while holding other variables’ constant show that the mean cell size and the mean cell wall thickness decreased, while the cell density and the open porosity increased with increase of pressure. Tensile modulus of PLA foams decreased with increasing the saturation pressure, but the specific tensile modulus of PLA foams was still 15–80% higher than that of solid PLA. Tensile strength and elongation at break first increased with increasing saturation pressure up to 16 MPa and then decreased with further increasing saturation pressure (20 MPa and 24 MPa) at which opened-cell structure produced. Compressive modulus, compressive strength, and compressive yield stress also followed the same variation trend. The results indicated that not only cell size plays an important role in properties of PLA foams but also cell morphology can influence these properties significantly.

scholarly journals The malignant cells in a Lennert's lymphoma are T lymphocytes with a mature helper surface phenotype. A multiparameter flow cytometric analysis

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 426-429 ◽  
Author(s):  
KJ Stonesifer ◽  
NA Benson ◽  
SE Ryden ◽  
DF Pawliger ◽  
RC Braylan
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Malignant Neoplasm ◽  
Surface Antigens ◽  
Light Scatter ◽  
Malignant Cells ◽  
T Helper ◽  
T Helper Lymphocytes ◽  
Flow Cytometric ◽  
Multiparameter Analysis

Abstract The flow cytometric analysis of DNA content in cells obtained from a case of Lennert's lymphoma demonstrated the presence of a discrete hypotetraploid cell population. Correlated multiparameter analysis of DNA, light scatter, and surface antigens by flow cytometry showed that the hypotetraploid cells were intermediate to large cells expressing T11, T3, and T4 antigens and lacking B1 and T8 antigens. These findings suggest that Lennert's lymphoma represents a malignant neoplasm of T- helper lymphocytes.

Flow Cytometric Analysis of Lymphomas: Current Status and Usefulness

2006 ◽  
Vol 130 (12) ◽  
pp. 1850-1858
Author(s):  
Zahid Kaleem
Keyword(s):  
Flow Cytometry ◽  
Hodgkin Lymphoma ◽  
Flow Cytometric Analysis ◽  
Molecular Diagnostic ◽  
Routine Practice ◽  
Becton Dickinson ◽  
Color Flow ◽  
Non Hodgkin Lymphoma ◽  
Flow Cytometric ◽  
Diagnosis And Classification

Abstract Context.—Immunophenotyping has become a routine practice in the diagnosis and classification of most cases of non-Hodgkin lymphoma, and flow cytometry is often the method of choice in many laboratories. The role that flow cytometry plays, however, extends beyond just diagnosis and classification. Objective.—To review and evaluate the current roles of flow cytometry in non-Hodgkin lymphoma, to compare it with immunohistochemistry, and to discuss its potential future applications in the molecular diagnostic era. Data Sources.—The information contained herein is derived from peer-reviewed articles on the subject published in the English-language medical literature during the years 1980 to 2005 that were identified using PubMed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi, 1980–2005) search, various books and other sources on flow cytometry, and the author's personal experience of more than 10 years with flow cytometric analysis of lymphomas and leukemia using Becton-Dickinson (San Jose, Calif) and Beckman-Coulter (Miami, Fla) flow cytometers. Study Selection.—Studies were selected based on adequate material and methods, statistically significant results, and adequate clinical follow-up. Data Extraction.—The data from various sources were compared when the methods used were the same or similar and appropriate controls were included. Most of the studies employed 2-color, 3-color, or 4-color flow cytometers with antibodies from Becton-Dickinson, Beckman-Coulter, or DakoCytomation (Carpinteria, Calif). Results were evaluated from studies utilizing the same or similar techniques and flow cytometers. Only objective data analyses from relevant and useful publications were included for reporting and discussion. Data Synthesis.—Flow cytometry serves a variety of roles in the field of lymphoma/leukemia including rapid diagnosis, proper classification, staging, minimal residual disease detection, central nervous system lymphoma detection, evaluation of prognostic markers, detection of target molecules for therapies, ploidy analysis of lymphoma cell DNA, and evaluation of multidrug-resistance markers. It offers many advantages in comparison to immunohistochemistry for the same roles and provides uses that are either not possible or not preferable by immunohistochemistry such as multiparameter evaluation of single cells and detection of clonality in T cells. Conclusions.—By virtue of its ability to evaluate not only surface but also cytoplasmic and nuclear antigens, flow cytometry continues to enjoy widespread use in various capacities in lymphoma evaluation and treatment. Additional roles for flow cytometry are likely to be invented in the future and should provide distinctive uses in the molecular era.

Clinical Predictors of Abnormal Peripheral Blood Lymphocytoses Diagnosed by Flow Cytometry: An Algorithmic Approach That Can Be Applied in Routine Clinical Practice.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3932-3932
Author(s):  
Jared M. Andrews ◽  
Mitchel T. Holm ◽  
Jerome B. Myers
Keyword(s):  
Flow Cytometry ◽  
Peripheral Blood ◽  
Lymphocyte Count ◽  
Flow Cytometric Analysis ◽  
Clinical History ◽  
Lymphocytic Leukemia ◽  
Western World ◽  
Abnormal Phenotype ◽  
Flow Cytometric ◽  
The Mean

Abstract Background Elevated peripheral blood lymphocyte counts in adults can occur in benign reactive conditions as well as malignant disease processes. Chronic lymphocytic leukemia (CLL) is the most common adult hematologic malignancy of the western world affecting the middle aged and elderly. Less commonly B, T, and Natural Killer (NK) cell leukemia / lymphomas may also present with lymphocytosis. Flow cytometry has greatly improved the ability to detect low levels of abnormal lymphocyte populations in peripheral blood. It is, however, a relatively expensive test and clinical guidelines for its appropriate usage are not well defined. Methods We conducted a retrospective review of peripheral blood lymphocytoses that were submitted for flow cytometric analysis at Madigan Army Medical Center, Tacoma, WA from 2002 – 2004. Under laboratory protocol, all patients ≥ 50 years of age with an absolute lymphocyte count (ALC) of > 4 X 109 Cells/L had a peripheral smear evaluated by both a hematology technician and pathologist. Specimens determined to warrant flow cytometric analysis based on review of clinical history, prior lab values, degree of lymphocytosis, and morphology were either recommended for flow cytometry in a comment; or sent directly for analysis with the clinician’s approval. We reviewed complete blood counts (CBCs), previous flow cytometry results, as well as bone marrow and electronic clinical history. All patients with previous diagnoses of lymphoproliferative disorders (LPDs) or ALC < 4 X 109 Cells/L were excluded. Results Approximately 7,300 CBC specimens/month (3,400 from patients ≥ 50 years of age) were performed. Of these, an average of 44 specimens/month had a lymphocytosis of ≥ 4 X 109 Cells/L, from approximately 28 different patients. From this group 71 flow cytometric cases (an average of 2/month) were performed over the 2 year period. 42 cases (59%) had an abnormal phenotype. 27 had a phenotype consistent with CLL, and the other 15 were a mixture of LPDs involving B and T-lymphocytes as well as NK cells. Comparing normal phenotype to abnormal phenotype showed statistically significant differences between the mean age (n-60.4 ±7.5, abn-69.8±8.7), ALC (n-4.9±0.8, abn-9.2±8.1), and relative lymphocyte count (RLC) (n-43.9±7.5%, abn-59.3±8.8%). Conclusion Absolute lymphocyte counts ≥ 4 X 109 Cells/L in adults ≥ 50 years of age represent approximately 1% of the CBCs performed in our laboratory. Review of these cases by a pathologist is logistically feasible due to the low incidence. Our method of reviewing for morphology, clinical history, and past lymphocyte counts with comments to the ordering clinician yielded a high incidence of abnormal phenotype diagnoses when evaluated by flow cytometric analysis (59%). Age, ALC, and relative lymphocyte counts are variables that can be used to develop guidelines for determining the appropriateness of flow cytometric analysis. Patients < 52.4 years of age fall below two standards of deviation from the mean age of the abnormal phenotype group. The standard of deviation for mean ALC is very small (4.9±0.8), which indicates that counts > two standards of deviation above the mean, or 6.5 X 109 Cells/L, would correlate strongly with an abnormal phenotype. The same conclusion could be made with a RLC > 58.9%. In conclusion, patients ≥ 50 years of age with an ALC > 6.5 X 109 Cells/L or a RLC > 58.9% are likely to have a lymphoproliferative disorder and flow cytometric analysis is indicated.

scholarly journals The malignant cells in a Lennert's lymphoma are T lymphocytes with a mature helper surface phenotype. A multiparameter flow cytometric analysis

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 426-429 ◽  
Author(s):  
KJ Stonesifer ◽  
NA Benson ◽  
SE Ryden ◽  
DF Pawliger ◽  
RC Braylan
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometric Analysis ◽  
Malignant Neoplasm ◽  
Surface Antigens ◽  
Light Scatter ◽  
Malignant Cells ◽  
T Helper ◽  
T Helper Lymphocytes ◽  
Flow Cytometric ◽  
Multiparameter Analysis

The flow cytometric analysis of DNA content in cells obtained from a case of Lennert's lymphoma demonstrated the presence of a discrete hypotetraploid cell population. Correlated multiparameter analysis of DNA, light scatter, and surface antigens by flow cytometry showed that the hypotetraploid cells were intermediate to large cells expressing T11, T3, and T4 antigens and lacking B1 and T8 antigens. These findings suggest that Lennert's lymphoma represents a malignant neoplasm of T- helper lymphocytes.

scholarly journals Analysis of phorbol ester stimulated human megakaryocyte development

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 202-207
Author(s):  
BJ Roth ◽  
GW Jr Sledge ◽  
JE Straneva ◽  
J Brandt ◽  
M Goheen ◽  
...  
Keyword(s):  
Phorbol Esters ◽  
Surface Membrane ◽  
Flow Cytometric Analysis ◽  
Leukemia Cell ◽  
Color Flow ◽  
Flow Cytometric ◽  
Differentiating Agents ◽  
Membrane Antigens ◽  
Cytoplasmic Maturation

Megakaryocytes are relatively rare components of human bone marrow, making the study of their maturation difficult. Phorbol esters can act as differentiating agents in a number of cell systems including murine megakaryocytes. We report the effects of phorbol esters on the previously described long-term human megakaryocytic leukemia cell culture, EST-IU. While two nontransforming phorbols fail to affect these cells, the transforming phorbol 12-O-tetradecanoylphorbol-13- acetate (TPA) induces a phenotype with characteristics of more mature megakaryocytes in a dose-related manner. This phenotype includes an increased adherence to untreated plastic or glass, polyploidization, an increase in cell size, and increased expression of both platelet glycoproteins and factor VIII-related antigen. Two-color flow cytometric analysis allowed simultaneous determinations of DNA content and the expression of surface membrane antigens or alpha-granule constituents, providing evidence that nuclear, membrane, and cytoplasmic maturation occur in parallel in this cellular system. TPA- induced maturation of EST-IU cells provides an important new cellular model for the further study of human megakaryocyte development.

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