Manufacturing process for fresh and frozen stem cells

摘要

The present application discloses a method of obtaining multi-lineage stem cells or progenitor cells by allowing a sample of cells to settle in a container; transferring supernatant from the container to another container; and eventually isolating a colony from the supernatant after several transfer/settle processes and expanding further, and optionally freezing the cells thus obtained.

主权项

1. A method of isolating multi-lineage stem cells or progenitor cells, comprising:
(i) allowing a sample of cells to settle in a first container without centrifugation, wherein the denser cells settle to the bottom of the container and comparatively less dense cells are present in the supernatant liquid; (ii) transferring the supernatant liquid that contains the less dense cells from the first container to a second container; and (iii) allowing the cells that are present in the supernatant liquid to settle without centrifugation, wherein the denser cells settle to the bottom and comparatively less dense cells are present in the supernatant liquid; (iv) repeating steps (ii) and (iii) at least three times; (v) isolating a single colony of multi-lineage stem cells or progenitor cells from the bottom of the container; (vi) transferring cells from the colony into a growth media and growing the cells; (vii) expanding the cells from 4 to 14 passages; and (viii) obtaining a bank of multi-lineage stem cells or progenitor cells.