Quantification of Osteoclasts in Culture, Powered by Machine Learning

Frontiers in Cell and Developmental Biology

Ayelet Bergman¹  Shahar Eshed¹  Yishay Mansour¹  Amir Globerson¹  Edo Cohen-Karlik¹  Omer Nestor¹  Yankel Gabet²  Zamzam Awida³  Sapir Ben Yosef³  Drorit Neumann³  Michelle Kadashev³  Hussam Saed³ 

[1] Blavatnik School of Computer Science, Tel Aviv University, Tel Aviv, Israel;Department of Anatomy and Anthropology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel;Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel;

关键词: osteoclasts;    automatic quantification of osteoclasts;    machine learning;    object detection;    deep learning;    convolutional neural network (CNN);   

DOI  :  10.3389/fcell.2021.674710

来源: DOAJ

【 摘 要 】

In vitro osteoclastogenesis is a central assay in bone biology to study the effect of genetic and pharmacologic cues on the differentiation of bone resorbing osteoclasts. To date, identification of TRAP+ multinucleated cells and measurements of osteoclast number and surface rely on a manual tracing requiring specially trained lab personnel. This task is tedious, time-consuming, and prone to operator bias. Here, we propose to replace this laborious manual task with a completely automatic process using algorithms developed for computer vision. To this end, we manually annotated full cultures by contouring each cell, and trained a machine learning algorithm to detect and classify cells into preosteoclast (TRAP+ cells with 1–2 nuclei), osteoclast type I (cells with more than 3 nuclei and less than 15 nuclei), and osteoclast type II (cells with more than 15 nuclei). The training usually requires thousands of annotated samples and we developed an approach to minimize this requirement. Our novel strategy was to train the algorithm by working at “patch-level” instead of on the full culture, thus amplifying by >20-fold the number of patches to train on. To assess the accuracy of our algorithm, we asked whether our model measures osteoclast number and area at least as well as any two trained human annotators. The results indicated that for osteoclast type I cells, our new model achieves a Pearson correlation (r) of 0.916 to 0.951 with human annotators in the estimation of osteoclast number, and 0.773 to 0.879 for estimating the osteoclast area. Because the correlation between 3 different trained annotators ranged between 0.948 and 0.958 for the cell count and between 0.915 and 0.936 for the area, we can conclude that our trained model is in good agreement with trained lab personnel, with a correlation that is similar to inter-annotator correlation. Automation of osteoclast culture quantification is a useful labor-saving and unbiased technique, and we suggest that a similar machine-learning approach may prove beneficial for other morphometrical analyses.

【 授权许可】

Unknown