Mechanotransduction by the synergistic action of heterotypic cell interactions
The candidates are screened and then employed by administering to patients in need thereof of a drug candidate that affects heterotypic intercellular mechanotransduction. At least two types of cells are labeled with distinct intracellular fluorescent marker labels and combined to form a cell suspension and cultured to form a microtissue, such as spheroids. The cells or the spheroids are combined with a drug candidate, either before, during or after forming the spheroids. The distribution of the different cell types is compared to that of essentially the same suspension culture in the absence of the drug candidate. Alternatively, the cell power of cells cultured in a non-adherent mold that determines at least in part, the shape of microtissue formed is measured and compared with essentially the same cell suspension cultured in the same manner in the absence of a drug candidate. A patient in need thereof can be administered a drug identified as affecting heterotypic intercellular mechanotransduction. Alternatively, a macrotissue can be formed that is employed in therapeutic treatment of a patient in need thereof.
1. A method for screening drug candidates that affect heterotypic intercellular mechanotransduction, comprising the steps of:
a) labeling at least two types of cells with distinct intracellular fluorescent marker labels;
b) seeding a cell culture medium with the at least two types of cells in a suspension;
c) combining a drug candidate with the cells in suspension, and then culturing the cells in a non-adherent mold to thereby form spheroids, or culturing the cells in the non-adherent mold to form spheroids and then combining the spheroids with the drug candidate, or culturing the cells in the non-adherent mold to form spheroids while combining the cells with the drug candidate; and
d) comparing the distribution of the at least two different types of cells to that of spheroids of essentially the same cell suspension cultured in the absence of the drug candidate.
2. The method of claim 1 , wherein at least one of the types of cells is a connective tissue cell.
3. The method of claim 2 , wherein the connective tissue is a fibroblast cell.
4. The method of claim 2 , wherein the connective tissue is a myofibroblast cell.
5. The method of claim 1 , wherein at least one of the types of cells is a parenchymal cell.
6. The method of claim 5 , wherein the parenchymal cell is at least one member selected from the group consisting of an epithelial cell, a muscle cell, a neural cell, kidney cells, lung cells, cardiomyocyte cells and liver cells.