Microcarrier for cell culture and method for preparing the same
The present application relates to microcarrier particles for cell culture, a method for preparing the particles, and a cell culture medium composition including the particles. According to the present application, a microcarrier having a high degree of uniformity in shape or form, having porosity, and advantageous for cell attachment and isolation of cultured cells is provided.
1 . A method for preparing microcarrier particles for cell culture, the method comprising the steps of:
mixing a continuous phase composition (A) satisfying the following conditions 1 and 2; and a dispersive phase composition (B) containing a polymerizable monomer (b1) and then performing suspension polymerization,
wherein the dispersive phase composition (B) comprises a hydrocarbon oil (b3) in an amount of 10 to 20% by weight based on the total content of the dispersive phase composition, wherein the microcarrier particles include microcarrier particles having a density in the range of 0.95 g/cm 3 to 1.00 g/cm 3 :
45 mN/m<Surface Tension of Continuous Phase Composition≤54 mN/m [Condition 1]
Viscosity of Continuous Phase Composition≥2.0 cp, and [Condition 2]
wherein, in the [Condition 1], the surface tension is measured at a normal temperature according to a ring method, and in the [Condition 2], the viscosity is measured under conditions of a shear rate in the range of 66 to 264 l/s and a normal temperature.
2 . The method for preparing microcarrier particles for cell culture according to claim 1 , wherein the continuous phase composition (A) comprises water and poly(vinyl alcohol) (PVA).
3 . The method for preparing microcarrier particles for cell culture according to claim 2 , wherein the polyvinyl alcohol (PVA) has a weight average molecular weight in the range of 80,000 to 190,000 and a hydrolyzed degree in the range of 80 to 99%.
4 . The method for preparing microcarrier particles for cell culture according to claim 2 , wherein the continuous phase composition (A) contains at least 1.0% by weight of the polyvinyl alcohol based on the total weight of the continuous phase composition.
5 . The method for preparing microcarrier particles for cell culture according to claim 1 , wherein the dispersive phase composition (B) comprises a styrene monomer as the polymerizable monomer (b1).
6 . The method for preparing microcarrier particles for cell culture according to claim 5 , wherein the dispersive phase composition (B) further comprises a crosslinking agent (b2).
7 . The method for preparing microcarrier particles for cell culture according to claim 6 , wherein the dispersive phase composition (B) contains the crosslinking agent in an amount of 3 to 300 parts by weight based on 100 parts by weight of the styrene monomer.
8 . The method for preparing microcarrier particles for cell culture according to claim 1 , wherein the hydrocarbon oil has a density in the range of 0.750 g/cm 3 to 0.800 g/cm 3 .
9 . The method for preparing microcarrier particles for cell culture according to claim 1 , wherein the hydrocarbon oil contains a linear or branched saturated hydrocarbon compound having 12 to 50 carbon atoms.
10 . The method for preparing microcarrier particles for cell culture according to claim 1 , wherein the suspension polymerization is performed under conditions of a temperature of 80 to 95° C. and at a speed of 300 to 900 rpm.
11 . The method for preparing microcarrier particles for cell culture according to claim 1 , wherein
the step of mixing the continuous phase composition (A) and the dispersive phase composition (B) followed by applying a shearing force to homogenize the dispersive phase composition (B) in the continuous phase composition (A) in the form of droplets; and
the dispersive phase composition (B) is subjected to the suspension polymerization.
12 . The method for preparing microcarrier particles for cell culture according to claim 1 , further comprising
a step of adding an additional amount of the continuous phase composition (A) during the suspension polymerization when the polymerization rate related to the suspension polymerization is in the range of at least 5% to 70%.
13 . The method for preparing microcarrier particles for cell culture according to claim 1 ,
wherein at least 80% of the microcarrier particles have a single shape where satellite particles are not present on the surface of the particles.
14 . The method for preparing microcarrier particles for cell culture according to claim 1 ,
wherein the microcarrier particles include porous microcarrier particles that are spherical and have pores of 5 μm or less.
15 . The method for preparing microcarrier particles for cell culture according to claim 1 ,
wherein the microcarrier particles include microcarrier particles having a diameter in the range of 90 to 250 μm.