Systems, methods and apparatus for adaptive passage of a culture of cells
Apparatus, systems and methods for the adaptive passage of a culture of cells and apparatus and methods for dissociating cell colonies are described. The systems may include an imaging module, a pipette module, a handling module, and/or a stage module. Coordinated operation of the modules, optionally in an automated manner, is effected by at least one processor based on one or more characteristics of the culture of cells calculated from one or more images captured at more than one time point. A first apparatus for adaptive passage of a culture cells includes an imaging module and at least one processor, which apparatus may be included in the systems or used in the methods. A second apparatus for dissociating cell colonies, may also be included in the systems or used in the methods, includes impact bumper(s) collidable with impact bracket(s) to transmit a dissociative force to a culture of cells.
1. A method for adaptive passage of one or more culture of cells, the method comprising:
capturing by an imaging module comprising a camera one or more dark-field images of the one or more culture of cells at a first time point and at one or more subsequent time points;
calculating by at least one processor one or more characteristics of the one or more culture of cells, based on the one or more images; and
outputting an adaptive passaging protocol based on the calculated one or more characteristics of the one or more culture of cells, the adaptive passaging protocol providing a split ratio and/or a passaging time for each of the one or more culture of cells to reach a threshold level of the one or more characteristics from the first time point to the one or more subsequent time points in a subsequent passage;
wherein the adaptive passaging protocol provides parameters comprising the split ratio and/or the passaging time for each of the one or more culture of cells to reach the threshold level of the one or more characteristics from the first time point to the one or more subsequent time points in the subsequent passage using a regression model to output the parameters.
2. The method of claim 1 , wherein the one or more characteristics are compared against corresponding one or more characteristics of a control culture or a standard.
3. The method of claim 1 , wherein the one or more characteristics includes:
a) a measure of a confluence of the culture of cells;
b) a measure of a morphology of cells or colonies of the culture of cells;
c) a measure of differentiation of cells or colonies of the culture of cells;
d) a measure of colony size distribution of the culture of cells;
e) a measure of the change of a), b), c), or d) from the first time point to the one or more subsequent time points; or
f) a measure of a) relative to b), c) or d), a measure of b) relative to a), c), or d), a measure of c) relative to a), b), or d), or a measure of d) relative a), b), or c).
4. The method of claim 3 , wherein the threshold level is:
i. for a), between 30-90% for cell or colony confluence;
ii. for b), between ±30% of a control culture;
iii. for c), between 0 to 30% of a control culture in a maintenance protocol or between 50% to 100% of a control culture in a differentiation protocol; or
iv. for d), within 15% of a mean colony size distribution of a control culture, or a subfraction thereof.
5. The method of claim 1 , wherein the one or more characteristics calculated in respect of the culture of cells and a second culture of cells in a second cell culture vessel is different at the first time point or at the one or more subsequent time points and the one or more characteristics are more consistent in the subsequent passage.
6. The method of claim 1 , further comprising obtaining a suspension of cells from the culture of cells and seeding some or all of the suspension of cells in a daughter cell culture vessel.
7. The method of claim 6 , wherein obtaining the suspension of cells includes aspirating the cell culture medium from the cell culture vessel and contacting the culture of cells in the cell culture vessel with a detachment solution.
8. The method of claim 7 , wherein the detachment solution is a fractionation solution, and the fractionation solution selectively detaches either a first population of differentiated cells or a second population of undifferentiated cells from a wall of the cell culture vessel.
9. The method of claim 1 , wherein the culture of cells are pluripotent stem cells, optionally human pluripotent stem cells.
10. The method of claim 1 , wherein the method is carried out using an apparatus, the apparatus comprising:
an imaging module for capturing one or more dark-field images of the one or more culture of cells at a first time point and at one or more subsequent time points; and
at least one processor communicatively coupled to the imaging module, the processor configured to:
receive from the imaging module the one or more images of the one or more culture of cells at a first time point and at one or more subsequent time points;
calculate one or more characteristics of the one or more culture of cells, based on the one or more images received from the imaging module; and
output the adaptive passaging protocol;
wherein the imaging module includes a camera capable of resolving a well of a culture dish, a colony of cells within the well, or a single cell in the well.
11. The method of claim 10 , wherein the method is carried out using a system, the system comprising the apparatus, wherein the at least one processor is also communicatively coupled to one or more of:
a pipette module having one or more pipettes for drawing a fluid from a cell culture vessel through a pipette tip mateable with an end of the one or more pipettes;
a liquid dispenser module spaced apart from the pipette module, the liquid dispenser module in fluid communication with more than one solution reservoir; and
a handling module having a pair of opposable arms for gripping and transporting the cell culture vessel or a lid thereof or a daughter cell culture vessel or a lid thereof within the system,
wherein the at least one processor coordinates operation of the apparatus and one or more of the pipette module, the liquid dispenser module, and the handling module, and wherein the liquid dispensing module includes more than one conduits and each conduit is in fluid communication with a separate one of the more than one solution reservoirs.
12. The method of claim 11 , wherein the system is automated.