Method for producing megakaryocytes, platelets and/or thrombopoietin using mesenchymal cells

Patent metadata

Field	Value
Publication number	US-2016177265-A1
Application number	US-201414899828-A
Country	US
Kind code	A1
Filing date	Jun 27, 2014
Priority date	Jun 28, 2013
Publication date	Jun 23, 2016
Grant date	—

Abstract

Provided is a megakaryocyte and/or platelet production method, enabling to produce a megakaryocyte and/or platelet from mesenchymal cells such as preadipocytes in a relatively short period of time, simply, in a large amount and at lower cost or more efficiently in vitro and a method for producing TPO simply and in a larger amount. A first invention is a method for producing a megakaryocyte and/or platelet, comprising culturing a mesenchymal cell in a mesenchymal cell culturing basic medium containing an iron ion and an iron transporter and collecting megakaryocytes and/or platelets from a culture. A second invention is a method for producing thrombopoietin, comprising culturing a mesenchymal cell or mesenchymal cell-derived megakaryocyte in a mesenchymal cell culturing basic medium containing an iron ion and an iron transporter and collecting thrombopoietin from a culture. A third invention is a method for producing thrombopoietin, comprising culturing a preadipocyte in a preadipocyte culturing basic medium containing dexamethasone, 3-isobutyl-1-methylxanthine and insulin and collecting thrombopoietin from a culture.

First claim

1 . A method for producing a megakaryocyte and/or platelet, comprising culturing a mesenchymal cell in a mesenchymal cell culturing basic medium containing an iron ion and an iron transporter, and collecting the megakaryocyte and/or platelet from a culture.

2 . (canceled)

3 . The production method according to claim 1 , wherein the mesenchymal cell is a CD31 negative and CD71 positive mesenchymal cell.

4 . The production method according to claim 1 , wherein the CD31 negative and CD71 positive mesenchymal cell is further positive to c-MPL.

5 . The production method according to claim 1 , wherein the iron transporter is transferrin.

6 . The production method according to claim 1 , wherein the iron ion and iron transporter are an iron-bound transferrin.

7 . The production method according to claim 1 , wherein the culturing is performed for 5 to 17 days.

8 . (canceled)

9 . The production method according to claim 1 , wherein a yield of the megakaryocyte and/or platelet is 270 to 1,080%.

10 . The production method according to claim 1 , wherein a percentage of platelets contributing to thrombus formation as measured by thrombus formation analysis of the platelets is 70% to 85%.

11 . (canceled)

12 . The production method according to claim 1 , further comprising a step of purifying the megakaryocyte and/or platelet from the culture.

13 . (canceled)

14 . A megakaryocyte and/or platelet produced by the production method according to claim 1 .

15 . (canceled)

16 . A method for constructing a megakaryocyte and/or platelet bank, comprising the following steps a) to c):
a) providing various types of mesenchymal cells of different HLA types and/or HPA types; b) selecting a specific HLA type and/or HPA type of mesenchymal cells among the mesenchymal cells provided in the step a); and c) preparing the specific HLA type and/or HPA type of megakaryocyte and/or platelet using the mesenchymal cells selected in the step b) by the production method according to claim 1 .

17 .- 18 . (canceled)

19 . The production method according to claim 3 , wherein the CD31 negative and CD71 positive mesenchymal cell is further positive for c-MPL.

20 . The production method according to claim 3 , wherein the iron transporter is transferrin.

21 . The production method according to claim 4 , wherein the iron transporter is transferrin.

22 . The production method according to claim 19 , wherein the iron transporter is transferrin.

23 . The production method according to claim 3 , wherein the iron ion and iron transporter are an iron-bound transferrin.

24 . The production method according to claim 4 , wherein the iron ion and iron transporter are an iron-bound transferrin.

25 . The production method according to claim 19 , wherein the iron ion and iron transporter are an iron-bound transferrin.

26 . The production method according to claim 3 , wherein the culturing is performed for 5 to 17 days.

27 . The production method according to claim 4 , wherein the culturing is performed for 5 to 17 days.

Assignees

• Univ Keio
• Kanagawa Kagaku Gijutsu Akad

Inventors

• Matsubara Yumiko
• Zama Takeru
• Ikeda Yasuo
• Uruga Yukako
• Suda Toshio
• Matsuoka Sahoko

Classifications

• C12N2511/00

  Cells for large scale production · CPC title

• C12N2506/1384

  from adipose-derived stem cells [ADSC], from adipose stromal stem cells · CPC title

• C12N2501/145

  Thrombopoietin [TPO] · CPC title

• C07K14/61

  Growth hormone [GH], i.e. somatotropin · CPC title

• C12N5/0644 Primary

  Platelets; Megakaryocytes · CPC title