Modern biomaterial biocompatibility research focuses the biomaterial hierarchic effects on multipotent mesenchymal stromal cell (MMSC) behavior (that occur at the nano-, micro- and macroscales) because MMSCs are the fundamental units that produce/regenerate bone tissue. Leukemia initiation and progression are connected with a disfunction of health cell microenvironment and MMSCs. Continuous monitoring of MMSC and tumor cell interaction is a promising tool for oncology, cellular biology, biotechnology and environmental research. The aim was to investigate a modulation of in vitro interaction of human MMSCs and leukemic T lymphoblast-like cells (Jurkat T cells) caused by micro-arc multilevel calcium phosphate (CP) coating with the help of Cell-IQ and RTCA advanced tools for continuous monitoring. An average velocity of cell division (AVCD) of human adipose-derived MMSCs (hAMMSCs) contacted in vitro with allogenic Jurkat line of human leukemic T lymphoblasts (Jurkat T cells) was studied by means of Cell-IQ v2 MLF integrated phase-contrast microscopic platform for real-time surveillance imaging of living cells. Both 50 μL suspensions (5×104 viable karyocytes) of the CD73CD90CD105+ adherent cells and Jurkat T cells were applied into the center of the well of 12-well plastic plates for 7 days at 100% humidity in a 5% CO2 atmosphere at 37°C until a monolayer formation. A nutrient medium was once replaced. To determine cell invasion (chemotactic motility) through 8 μm pores the real-time cell analysis (RTCA DP Analyzer) with the CIM-plate was used. AVCD of fibroblast-like adherent hAMMSCs was 0.27-0.63 divisions/h. CP coating diminished significantly the percent of dividing hAMMSCs contacted with leukemic Jurkat T cells. RTCA system showed significant hAMMSC invasion towards tumor cells and not vice versa. For all this, cellular interaction led to increasing viability of Jurkat T cells and decreasing hAMMSC viability. Thus, tumor Jurkat T cells could control a fate of health hAMMSCs and promote stromal microenvironment for survivability of tumor clones by means of secretable molecular products. Multilevel micro-arc CP coating forms bone-like inorganic structure that is capable to modulate in vitro interaction of human MMSCs and leukemic T lymphoblasts. The results obtained may be useful for replacement surgery applications of orthopedic implants in cancer patients.