A review of state-of-the-art stereology for better quantitative 3D morphology in cardiac research

Mühlfeld, Christian; Nyengaard, Jens Randel; Mayhew, Terry M.

doi:10.1016/j.carpath.2008.10.015

Cited by 139 publications

(126 citation statements)

References 97 publications

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“…Cardiomyocyte silhouettes were measured on perpendicular sections of the papillary muscles (28), and the results led researchers to conclude that cardiomyocyte enlargement is the sole mechanism for postnatal growth. By today's standards, this study was limited by selection, anisotropy, and counting biases (28,29). Although other reports suggested that cardiomyocytes in human infants show mitotic activity (30)(31)(32) and that the number of cardiomyocytes may double in the first year of life (33), the cell-static view has dominated most of our thinking about physiologic heart growth in humans.…”

Section: Discussionmentioning

confidence: 99%

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Cardiomyocyte proliferation contributes to heart growth in young humans

Mollova¹,

Bersell

Walsh³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

647

578

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The human heart is believed to grow by enlargement but not proliferation of cardiomyocytes (heart muscle cells) during postnatal development. However, recent studies have shown that cardiomyocyte proliferation is a mechanism of cardiac growth and regeneration in animals. Combined with evidence for cardiomyocyte turnover in adult humans, this suggests that cardiomyocyte proliferation may play an unrecognized role during the period of developmental heart growth between birth and adolescence. We tested this hypothesis by examining the cellular growth mechanisms of the left ventricle on a set of healthy hearts from humans aged 0-59 y (n = 36). The percentages of cardiomyocytes in mitosis and cytokinesis were highest in infants, decreasing to low levels by 20 y. Although cardiomyocyte mitosis was detectable throughout life, cardiomyocyte cytokinesis was not evident after 20 y. Between the first year and 20 y of life, the number of cardiomyocytes in the left ventricle increased 3.4-fold, which was consistent with our predictions based on measured cardiomyocyte cell cycle activity. Our findings show that cardiomyocyte proliferation contributes to developmental heart growth in young humans. This suggests that children and adolescents may be able to regenerate myocardium, that abnormal cardiomyocyte proliferation may be involved in myocardial diseases that affect this population, and that these diseases might be treatable through stimulation of cardiomyocyte proliferation.heart failure | pediatrics H eart failure, a leading public health problem worldwide (1), is linked to the loss of cardiomyocytes (2-4). The only currently available, definitive therapy-heart transplantation-is limited by donor availability. New approaches, such as cell transplantation, have shown encouraging results in clinical trials (5, 6). However, a third, complementary strategy has emerged, based on stimulating endogenous regenerative mechanisms. One approach for developing such regeneration strategies is to examine the cellular mechanisms of myocardial growth, since mechanisms of regeneration should be similar to the mechanisms of development.Although stem and progenitor cells are important for morphogenesis of the myocardium, developmental growth in a number of nonhuman species is largely driven by cardiomyocyte proliferation (7-9). In biological models that, unlike adult humans, regenerate myocardium, cardiomyocyte proliferation is important for regeneration as well as postnatal heart growth (10, 11). For example, in mice, developmental cardiomyocyte proliferation continues for up to day 7 after birth, which coincides with the loss of regenerative capacity (11,12). The close temporal relationship between cardiomyocyte proliferation and heart regeneration in animals raises the question of whether and to what age and extent cardiomyocyte proliferation plays a role in humans. The answer may help us understand the endogenous regenerative potential of the human heart and possibly indicate strategies for stimulating cardiomyocyte proliferation to reg...

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Section: Discussionmentioning

confidence: 99%

“…Software analysis of the converted binary images was performed with Image Processing and Analysis in Java (Image J). To quantify the number of cardiomyocyte nuclei per cubed centimeter, we used the optical dissector method (29) and validated it with two cardiomyocyte-specific structural markers (SI Appendix, Fig. S3).…”

Section: Methodsmentioning

confidence: 99%

Cardiomyocyte proliferation contributes to heart growth in young humans

Mollova¹,

Bersell

Walsh³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

647

578

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“…Isotropic uniform random sections are necessary in order to estimate some stereological parameters including the oocyte's surface area [12][13][14][15]. The ovaries should be sectioned based on the 'isotropic Cavalieri' design to emphasize the fact that the sections are parallel and hence not independent and can be used in estimation of the ovary volume [13,15]. To obtain these sections, a method called 'isector' was applied.…”

Section: Methodsmentioning

confidence: 99%

“…Estimation of the total ovarian volume, 'V(Ref) ', was made according to the Cavalieri principle and using a stereomicroscope connected to a computer, at the final magnification of 20 ×. A brief description is presented under Figure 1 [13,15].…”

Section: Methodsmentioning

confidence: 99%

Stereological estimation of ovarian oocyte volume, surface area and number: application on mice treated with nandrolone decanoate

Karbalay‐Doust

Noorafshan

2012

Folia Histochem Cytobiol.

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Changes in the number and size of oocytes can lead to fertilization problems. The present study aimed to evaluate the number, volume, and surface area of oocytes in healthy as well as nandrolone decanoate-treated (ND) mice using stereological methods. Five control mice received vehicle, and five ND-treated mice received ND. Using the 'isotropic Cavalieri' design', the ovary was sectioned. The volume of the ovary (cortex and medulla) was estimated. The oocytes' volume and surface area were estimated using the invariator. The number of the oocytes was estimated using an optical disector. The volumes of the ovary, cortex, and medulla decreased ~50% in the ND-treated mice. The mean number (coefficient of variation) of preantral, antral, and atretic oocytes in the control ovary were 1,690 (0.29), 2,100 (0.52), and 3,900 (0.2), respectively, which decreased ~54%, ~87%, and ~91%, respectively in the ND-treated animals. The mean volume (coefficient of variation) of the preantral, antral, and atretic oocytes were 86,000 (0.27), 110,000 (0.48), and 27,000 (0.33) mm 3 , respectively. The mean surface area (coefficient of variation) of the three types of oocytes were 9,000 (0.24), 9,900 (0.28), and 4,700 (0.21) mm 2 , respectively. These parameters remained unchanged in the ND-treated mice. ND induces reduction in the number of oocytes, but not in the volume or the surface area.

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“…Design-based stereology can, however, provide fundamental information on the structural changes in the pathological heart that cannot be obtained using physiological, biochemical, or molecular techniques. Quantitative estimates of the number of nuclei and myocytes, length and surface area of capillaries can only be performed using design-based stereology (for review see Muhlfeld et al, 2009). Using quantitative microscopic methods without the necessary background, however, may introduce unpredictable errors leading to invalid biological interpretations and conclusions.…”

mentioning

confidence: 99%

The Application of Stereological Methods for Estimating Structural Parameters in the Human Heart

Tang

Nyengaard

Andersen

et al. 2009

The Anatomical Record

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This study describes and exemplifies generally applicable designbased stereological methods for obtaining quantitative estimates of the numbers and sizes of capillaries, cardiomyocytes, and cardiomyocyte nuclei in immersion-fixed human left ventricles (N ¼ 6). The design-based stereological methods are valid in all cardiac investigations onto quantifying changes in structure and function as seen under various conditions such as during development, aging, hypertrophy, and following ischemia/ reperfusion. The applied principles of unbiased stereology were as follows: 1) uniform random sampling was taken at all levels, also in respect to orientations, for estimates of length and mean sizes. 2) All global structural quantities were estimated as total quantity ¼ density Â volume of the left ventricle. As an example, the left ventricle contains 1.5 Â 10 9 capillaries with a total length of just below 200 km. 3) Stereological methods were used for estimating the volume density, surface area density, and length density of capillaries and cardiomyocytes. The numerical density of cardiomyocyte nuclei and capillaries was estimated, using the optical and physical disector, respectively. 4) In all local quantities, ''size'' was estimated either directly, using unbiased estimators to obtain the average individual size and size distribution parameters, or indirectly, using the relationship that: average size ¼ total quantity/total number. In the six hearts constituting this study, we observed the anticipated correlation between left ventricular volume and global estimates such as total number of capillaries. There were no correlation between local quantities and total left ventricular volume (e.g., average star volume of individual cardiomyocytes). Anat Rec, 292:1630Rec, 292: -1647Rec, 292: , 2009. V V C 2009 Wiley-Liss, Inc.

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A review of state-of-the-art stereology for better quantitative 3D morphology in cardiac research

Cited by 139 publications

References 97 publications

Cardiomyocyte proliferation contributes to heart growth in young humans

Cardiomyocyte proliferation contributes to heart growth in young humans

Stereological estimation of ovarian oocyte volume, surface area and number: application on mice treated with nandrolone decanoate

The Application of Stereological Methods for Estimating Structural Parameters in the Human Heart

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