US 12,234,478 B2
Method of autologous primary hair follicles preparation in 3D culture
Abhijit Bopardikar, Mumbai (IN); Oleksandr Kukharchuk, Mumbai (IN); Padma Priya Anand Baskaran, Mumbai (IN); Andrii Kukharchuk, Mumbai (IN); Sunil Pophale, Mumbai (IN); and Rohit R. Kulkarni, Mumbai (IN)
Assigned to REELABS PVT. LTD., Mumbai (IN)
Appl. No. 16/629,402
Filed by REELABS PVT. LTD., Mumbai (IN)
PCT Filed May 3, 2019, PCT No. PCT/IB2019/053629
§ 371(c)(1), (2) Date Jan. 8, 2020,
PCT Pub. No. WO2019/215557, PCT Pub. Date Nov. 14, 2019.
Claims priority of application No. 201821017071 (IN), filed on May 7, 2018.
Prior Publication US 2020/0149005 A1, May 14, 2020
Int. Cl. C12N 5/071 (2010.01); G01N 1/28 (2006.01)
CPC C12N 5/0627 (2013.01) [G01N 1/28 (2013.01); C12N 2500/00 (2013.01); C12N 2500/30 (2013.01); C12N 2501/11 (2013.01); C12N 2501/115 (2013.01); C12N 2501/117 (2013.01); C12N 2501/12 (2013.01); C12N 2501/125 (2013.01); C12N 2501/15 (2013.01); C12N 2501/165 (2013.01); C12N 2501/17 (2013.01); C12N 2501/25 (2013.01); C12N 2501/734 (2013.01); C12N 2501/998 (2013.01); C12N 2506/1346 (2013.01); C12N 2513/00 (2013.01); C12N 2533/56 (2013.01)] 10 Claims
 
1. A method for creating de novo autologous primary hair follicles in 3D culture by recreating the conditions of embryogenesis of primary hair follicles, which involves: isolating mesenchymal stem cells of donor hair follicles and their expansion; isolating of donor scalp skin cells and their temporary cryopreservation; seeding, using a sandwich method, mesenchymal stem cells of the donor's hair follicles and skin cells of the donor's scalp in a monolayer of fibrin gel based on a nutrient medium containing growth factors of fetal scalp hair follicles; lysing of the fibrin gel monolayer with exogenous plasmin; isolating de novo-formed primary donor hair follicles from a lysed fibrin gel monolayer by self-precipitation; washing the de novo-formed primary hair follicles of the donor with phosphate buffer; transplanting of de novo-formed primary hair follicles to a donor.