Chapter 2 Blood cell morphology and development

2.3 ORIGIN AND DEVELO !EN" O# RED BLOOD CELL$

There are three types of cellular elements in the blood - erythrocytes ( red cell%), leukocytes &'h(te cell%), and thrombocytes (platelet%). Each has its own functions and differs clearly from the others. In health, destruction is balanced by production and the quantity of circulating cells remain remarkably constant. ost of the blood cells are formed in the bone marrow. !onsequently, e"amination of marrow specimens often pro#ides information of great #alue in the diagnosis and treatment of hematologic disorders.

2.3.* !+ELO OIE$I$,

The prefi" -myelo-. generally refers to all aspects of bone marrow acti#ity and is not limited to granulocytic elements alone. The only e"ceptions to this rule are the terms myeloblast, myelocyte, and metamyelocyte, which refer to specific stages in granulocyte maturation. !onsists chiefly of erythropoiesis, granulopoiesis, and thrombopoiesis. onocytes appear to be formed in the marrow as well as elsewhere. $ymphopoiesis occurs largely in e"tramedullary sites such as the spleen, thymus, and lymph nodes. There is little doubt that a primiti#e, pluripotential reticulum cell or %stem cell% can undergo differentiation into a specific blast cell. The marrow reticulum cells are those from which the differentiated blood cell precursors arise. The reticulum cells in a &right's stained preparation are relati#ely large &2/.3/ 0 (n d(ameter) with abundant, light blue, opaque cytoplasm. The cell borders are typically irregular. These cells ordinarily ha#e one to two nucleoli with the nucleus being round to o#al, with a finely to coarse reticulated chromatin pattern. Figure II.1 lists the steps in the de#elopment of the #arious blood cells from the marrow reticulum cell.

2.3.2 ER+"1RO OIE$I$,

The nonnucleated red cells in the peripheral blood are deri#ed from nucleated precursors in the bone marrow. (istinct morphological and functional stages can be recogni)ed in the de#elopment of an erythrocyte and ser#e as a useful outline of erythropoiesis. *s a group, all of the immature and mature erythrocytes in the body are designated the erythron. any names ha#e been used for the de#elopmental stages of an erythrocyte. The terminology used in this syllabus is preferred since accurate definitions are a#ailable.

2.3.3 !A"2RA"ION $E32ENCE

*s cells de#elop, their morphology changes. These morphologic alterations occur gradually+ therefore, many cells are in transition from one named cell type to another. *s a general rule, cells in transition are classified towards the more differentiated cell type. Thus, if there is a question whether a cell is a rubricyte or a metarubricyte, call it the latter. Immature cells are generally larger and become smaller as they mature. #(g0re II.2 illustrates the relationship of cell si)e to maturation and di#ision. The nuclei of the immature cells are relati#ely large in relation to the amount of cytoplasm and become smaller with maturity. The chromatin of the nucleus in immature cells is delicate, fine and stippled. *s the cell matures, the chromatin becomes coarse, clumped and compact. ,ucleoli are found in the nucleus of immature cells. #(g0re II.3 illustrates these maturation changes.

2.3.4 ER+"1ROC+"E !A"2RA"ION $E32ENCE, 2.3.4.* R05r(5la%t 2.3.4.2 ror05r(cyte 2.3.4.3 R05r(cyte 2.3.4.4 !etar05r(cyte 2.3.4.6 Ret(c0locyte or polychromatoph(l(c cell 2.3.4.7 Erythrocyte

#(g0re II.*

Figure II.1

"he Relat(on%h(p o8 Cell $(9e to !at0rat(on and D(v(%(on

FIGURE II.3
!orpholog(c Change% Occ0rr(ng D0r(ng Red Blood Cell !at0rat(on

R2BRIBLA$",

-irst recogni)able cell in the erythrogenic series. * large round cell which contains a large round nucleus with a thin rim of royal blue cytoplasm. The nuclear chromatin is delicate and stippled. The nucleus contains one to se#eral nucleoli. The cytoplasm is somewhat scanty and stains #ery basophilic.

ROR2BRIC+"E,

!ell is similar to the .ubriblast but smaller in si)e. ,ucleoli are usually absent, but remnants of the nucleoli may still be present. The chromatin material is somewhat coarser. * perinuclear clear )one may be obser#ed. The cytoplasm stains basophilic.

R2BRIC+"E,

!ell is smaller than the /rorubricyte. ,uclear chromatin material is coarsely clumped separated by light streaks gi#ing the so-called cartwheel appearance. ,ucleus is round and stains #ery dark. !ytoplasm stains #ery basophilic in the early rubricyte stage, but the blue color is diluted out by the pink color of hemoglobin as it matures toward the ne"t stages. This cell can be further subdi#ided according to the amount of hemoglobin in the cytoplasm into basophilic, polychromatophilic and normochromic rubricyte. The cell is the last cell in the erythrocytic series capable of cell di#ision under normal cond(t(on%. !ell di#ision stops when a critical hemoglobin concentration is reached. * deficiency of hemoglobin in the cell can result in e"tra di#isions and smaller cells. (E:A! LE: Iron deficiency results in microcytes.)

!E"AR2BRIC+"E,
The nucleus is small, pyknotic, and appears as a dark blue homogeneous mass without any distinct chromatin structure. The cytoplasm stains similarly to the mature erythrocyte. The cell is not found in the peripheral blood of normal healthy animals. &hen obser#ed in peripheral blood, it denotes a response to an anemic condition of at least 01-23 hours duration. The e"ception is the horse in that this species does not release nucleated red cells into the peripheral blood under any circumstances.

RE"IC2LOC+"E OR OL+C1RO!A"O 1ILIC CELL,

This cell is larger than the mature erythrocyte and is non-nucleated. !ell may contain a small round nuclear remnant called a 4owell-5olly body. !ytoplasm stains slightly basophilic with &right's stain. 4owe#er, when stained with a %0prav(tal.%ta(n such as new methylene blue or brilliant cresyl blue, precipitated ribrosomal .,* (reticulum) can be demonstrated within the cell. !ells are not found in health in the peripheral blood of the horse, cow, sheep, and goat. This means that the reticulocyte ripens in the bone marrow in these species. (ogs and cats may normally ha#e *.*.6; reticulocytes in the peripheral blood. The cell does not participate in normal rouleau" formation or pathologic agglutination+ is more resistant to crenation and lysis+ is less susceptible to mechanical trauma+ has a great e"cess of membrane in relation to its contents, but is able to synthesi)e hemoglobin.

ER+"1ROC+"E,

These are the mature non-nucleated red blood cells. !ell stains buff or reddish color.

2.3.6 CON"ROL O# ER+"1RO OIE$I$

Erythropoietin is accepted as the fundamental erythropoietic stimulus.It is produced by the kidney and6or li#er in most species.Its release is determined by tissue o"ygen le#els. Erythropoietin also causes an increased release of reticulocytes into the circulation &ret(c0locyto%(%). Endocrine glands such as the pituitary, thyroid, adrenals, and gonads affect erythropoiesis. These glands are responsible for general metabolic acti#ity and cellular o"ygen requirements.

2.4 ORIGIN AND DEVELO !EN" O# GRAN2LOC+"E$

7ranulocytes de#elop in the bone marrow from undifferentiated stem cells. 7ranulocytes are readily differentiated from the nucleated erythrocytes by their fine, reticulated chromatin structure and bluish cytoplasm.

2.4.* !A"2RA"ION $E32ENCE

aturation of the granulocytic series of cells is characteri)ed by the de#elopment of granules. 7ranules are initially formed in the progranulocytes and are called -pr(mary or a90roph(l- granules. These granules are pero"idase positi#e and de#elop from fusion of small proteinaceous material by the 7olgi comple". *s the maturation sequence continues, the primary or a)urophil granules lose their acid mucopolysaccharide and, therefore, will not stain with &right's stain. 4owe#er, the myelocyte stage is characteri)ed by the appearance of %specific% or secondary granules that persist throughout the maturation process. !ells which ha#e granules with an affinity for blue or basic dye are identified as basophils+ cells that are stained reddish-orange with the acid dye eosin are called eosinophils+ and the cells with granules which do not stain intensely with either dye are called neutrophils.

2.4.2 GRAN2LOC+"E !A"2RA"ION $E32ENCE

8e#eral distinct de#elopmental stages of granulocytes can be recogni)ed morphologically. The stages are9
2.4.2.* !yelo5la%t 2.4.2.2 rogran0locyte 2.4.2.3 !yelocyte .. Ba%oph(l< eo%(noph(l< ne0troph(l 2.4.2.6 !etamyelocyte .. Ba%oph(l< eo%(noph(l< ne0troph(l 2.4.2.7 Band Cell .. Ba%oph(l< eo%(noph(l< ne0troph(l

2.4.2.7 Segmented Cell -- Basophil eosinophil, neutrophil 2.4.2.* !+ELOBLA$"

The first cell that can be recogni)ed in the granulocytic series. It is the most immature granulocytic precursor. The cell possesses a relati#ely large, round to o#al nucleus, with one to se#eral nucleoli. The chromatin material of the nucleus is finely stippled or has a light ground glass appearance. The cytoplasm is somewhat scanty, basophilic, and does not contain granules.

2.4.2.2 ROGRAN2LOC+"E

The nuclear chromatin material is coarser and slightly more clumped than that of a myeloblast. .emnants of the nucleoli may still be present. The cytoplasm is less basophilic than the myeloblast and contains darkly stained non-specific granules called -pr(mary or a90roph(l(c gran0le%.% These granules are pero"idase positi#e.

2.4.2.3 !+ELOC+"E

The nucleus of the myelocyte remains somewhat round to o#al and the chromatin material is more closely clumped. This cell contains -%econdary- or specific granules that are identified by their staining properties as neutrophils, eosinophils, and basophils. These granules are pero"idase negati#e. Therefore, the myelocyte and all subsequent cells of the granulocytic series should be characteri)ed as neutrophils, eosinophils and basophils. These granules #ary greatly in shape, si)e and concentration in domestic species. *s maturation of the granulocytes continue, the -%pec(8(c gran0le%- will increase and the -a90roph(l(c gran0le%- will not take the &right's stain. The myelocyte stage is the last stage of cell di#ision and the first cell capable of phagocytosis.

2.4.2.4 !E"A!+ELOC+"E

This cell closely resembles the myelocyte. The nucleus is indented and often resembles a kidney bean. ,ucleoli are not present and the nuclear chromatin material is coarser and clumped. !ytoplasmic granules are also present.

2.4.2.6 BAND CELL

This cell has a horseshoe shaped nucleus. The opposite sides of the nucleus are more or less parallel. This cell may be differentiated from the metamyelocyte by the nuclear shape and the tendency for the nuclear sides to become parallel. The nuclear chromatin material is markedly clumped.

2.4.2.7 $EG!EN"ER CELL

The nucleus may be mono-lobed with clumped chromatin material, or may consist of se#eral lobes separated by constrictions or by filaments. The cytoplasm stains #ery faintly.

2.4.3 LE2=OC+"E L+$O$O!E$

The term lysosome is used to describe intracellular membranous sacs containing acid hydrolytic en)ymes. &hen leukocytes phagocyti)e, there is a release of lysosomal contents. /rimary granules (a90roph(l gran0le%) contain acid phosphatase, acid hydrolytic en)ymes, basic protein and one-third of the lyso)yme. 8econdary granules (%pec(8(c gran0le%) contain alkaline phosphatase, lactoferrin and two-thirds of the lyso)yme.

2.6 ORIGIN AND DEVELO !EN" O# AGRAN2LOC+"E$

The agranulocytic series is comprised of leukocytes de#oid of specific granulation. These cells generally originate in the lymphatic system, but like the granulocytic series, they may be produced elsewhere in the body. This series includes the lymphocytic and monocytic groups.

2.6.* L+! 1OC+"IC !A"2RA"ION $E32ENCE

The lymphocytic series refers to the de#elopment of the lymphocytes. The cells arise mainly from the reticular tissue of the lymph nodes and lymphoid tissue from which they deri#e their name. $ymphocytes are the most undifferentiated of all blood cells normally found in the peripheral blood. The nucleus does not become segmented and specific granules do not de#elop. Therefore, lymphocytes are -pero>(da%e negat(ve-. The lymphocyte cytoplasm does not de#elop past the blue stage. The stages in lymphocytic de#elopment are9

$ymphoblast /rolymphocyte $ymphocyte

2.6.*.* L+! 1OBLA$"

o o o o

!ell is similar to other blast cells. It is round or o#al, #ery large, with a large round to o#al reddish-purple nucleus. The nuclear chromatin material is fine and well distributed but perhaps more coarse than in myeloblasts. The nucleus contains one or two nucleoli. The cytoplasm is bluish and nongranular and forms a thin rim around the nucleus.

2.6.*.2 ROL+! 1OC+"E

o o o o

The nucleus is round or o#al in shape but smaller than the lymphoblast. The nuclear chromatin is coarse and slightly clumped. ,ucleoli or remnants of nucleoli may be present. There is an abundant amount of light blue cytoplasm around the nucleus. *lso, there may be a few a)urophilic granules in the cytoplasm.

2.6.*.3 L+! 1OC+"E

This is the mature cell of the lymphocytic series and the only cell form found in the peripheral blood in health. $ymphocytes #ary greatly in si)e and may be classified as small, medium or large. 4owe#er, si)e does not determine age of these cells. The cells are easily distorted and often appear in irregular shapes in stained preparations. The nuclear chromatin is condensed to form large, discrete almost solid clumps, with thickening of the nuclear membrane. ,ucleoli are absent. ,on specific granules may be obser#ed in the cytoplasm of these cells.

2.6.2 !ONOC+"IC !A"2RA"ION $E32ENCE

The monocytic series refer to the stages of de#elopment of the monocyte. These cells may be formed from .E cells. :ne of the most important sites of origin is the spleen. The stages in the monocytic de#elopment are9

onoblast /romonocyte onocyte

2.6.2.* !ONOBLA$"
The cell is large with a round or o#al nucleus. * nucleolus is present. The nuclear chromatin material is fine and well-distributed. There is a thin rim of clear blue cytoplasm around the nucleus. There are no granules present in the cytoplasm.

2.6.2.2 RO!ONOC+"E
The cell is somewhat smaller than the monoblast with the nucleus being irregularly-shaped.The nuclear chromatin material is fine and spongy. There may be a nucleolus or a remnant of the nucleolus present. The cytoplasm is grayish blue and may contain non-specific granules.

2.6.2.3 !ONOC+"E
The cell is larger than a neutrophil in the thin portions of a smear. The shape of monocytes is #ariable.The nuclei are usually round or kidney-shaped, but may be deeply indented or ha#e two or more lobes connected by narrow bands. ;lunt pseudopods and digesti#e #acuoles may be present. onocytes are most difficult to identify and to differentiate from other cells. They are frequently mistaken for immature neutrophils and large lymphocytes.The three most characteristic features of the monocytes and the most helpful in diagnosis are the dull grayish-blue color of the cytoplasm, blunt pseudopods and the brain-like con#olution of the nucleus.

2.7 ORIGIN AND DEVELO !EN" O# "hrom5ocyte%?platelet%

!ells of the megakaryocytic system are peculiar in that the nucleus undergoes multiple mitotic di#isions without cytoplasmic separation, thus producing giant polyploid cells. The multiple nuclei usually remain attached to each other and are often superimposed gi#ing a lobular appearance. The cytoplasm undergoes maturation changes characteri)ed by the de#elopment of granules and membranes, culminating in platelet differentiation and liberation. The stages in thrombocyte de#elopment are9

egakaryoblast /romegakaryocyte egakaryocyte Thrombocyte & latelet)

2.7.* !EGA=AR+OBLA$"
The cell is large, irregularly shaped with a single or se#eral round or o#al nuclei and with a blue, nongranular cytoplasm. ,ucleoli are usually present.

2.7.2 RO!EGA=AR+OC+"E
This cell differs from the megakaryoblast in that there are bluish granules in the cytoplasm ad<acent to the nucleus. The nucleus in this second stage of maturation has usually di#ided one or more times and the cell has increased in si)e.

2.7.3 !EGA=AR+OC+"E

The cell is #ery large with relati#ely large amounts of cytoplasm, and multiple nuclei. The cytoplasm contains numerous small, uniformly distributed granules that are reddish-blue in color.

2.7.4 "1RO!BOC+"E?platelet
/latelets are fragments of the cytoplasm of megakaryocytes. They #ary in si)e and shape from a barely #isible structure to masses larger than red cells or leukocytes. The cytoplasm stains a light blue and contains #ariable numbers of small blue granules (a90roph(l(c).