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US 12,540,309 B2Granted
Methods of isolating cells from placental tissue
AssigneeMeharry Medical College
InventorDonald J. Alcendor
Life of this patent
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Abstract
Methods for isolating a population of cells from tissue are disclosed herein. In particular, the present disclosure provides methods for isolating cytotrophoblasts from placental tissue. The methods include isolating a population of cells with the use of a laboratory apparatus that is coated with a lubricant, such as petroleum jelly. The methods described herein achieve consistent in vitro isolation of purified cells.
Claims15
A method for isolating and identifying a trophoblast cell, comprising:
- harvesting a placental villous tissue explant from a detached normal placenta;
- culturing the placental villous tissue explant in a culture medium under conditions suitable for proliferation of trophoblasts, wherein the culture medium comprises fetal bovine serum (FBS) and penicillin-streptomycin;
- cutting a tip and bulb portion off of a plastic pipette to expose a barrel portion of the pipette, wherein the barrel portion of the pipette has a first open end and a second open end;
- isolating, with the barrel portion of the pipette, the trophoblast cell by coating the first open end of the barrel portion with a lubricant, the lubricant selected from the group consisting of petroleum jelly, mineral oil, grease, and combinations thereof;
- placing the coated first open end of the barrel portion over the trophoblast cell to form an isolated trophoblast cell;
- treating the isolated trophoblast cell with forskolin;
- and
- staining the forskolin-treated trophoblast cell with an antibody to human chorionic gonadotropin (HCG) to verify identity.
5 more elements
A method for isolating and identifying a colony of trophoblast cells from placental tissue, comprising:
- obtaining a detached normal placenta;
- collecting a placental cotyledon from the detached normal placenta;
- harvesting a placental villous tissue explant from the placental cotyledon;
- culturing the placental villous tissue explant in a culture medium under conditions suitable for proliferation of trophoblasts, wherein the culture medium comprises fetal bovine serum (FBS) and penicillin-streptomycin;
- cutting a tip and bulb portion off of a plastic pipette to expose a barrel portion of the pipette, wherein the barrel portion of the pipette has a first open end and a second open end;
- isolating, with the barrel portion of the pipette, a colony of trophoblast cells by coating the first open end of the barrel portion with a lubricant, the lubricant selected from the group consisting of petroleum jelly, mineral oil, grease, and combinations thereof, and placing the coated first open end of the barrel portion over the colony of trophoblast cells to form an isolated colony;
- treating the isolated colony of trophoblast cells with forskolin;
- and
- staining the forskolin-treated trophoblast cells with an antibody to human chorionic gonadotropin (hCG) to verify identity.
6 more elements
Dependencies are inferred from the claim text.